MATERIALS AND METHODS FOR CONTROLLING REGULATION IN BIOSYNTHESIS IN SPECIES OF THE GENERA RALSTONIA OR CUPRIAVIDUS AND ORGANISMS RELATED THERETO

Abstract

Methods for increasing carbon-based chemical product yield in an organism by genetically modifying one or more genes involved in a stringent response and/or in a regulatory network, nonnaturally occurring organisms having increased carbon-based chemical product yield, and methods for use in production of carbon-based chemical products are provided.

Description

[0001] This patent application claims the benefit of priority from U.S. Provisional Patent Application Ser. No. 62/665,790 filed May 2, 2018, teachings of which are herein incorporated by reference in their entirety.

FIELD

[0002] The present invention relates to methods for increasing carbon-based chemical product yield in an organism by modifying one or more genes involved in a stringent response and/or in a regulatory network, nonnaturally occurring organisms having increased carbon-based chemical product yield, and methods for use in production of carbon-based chemical products.

BACKGROUND

[0003] Organisms have a limited ability to control their environment and therefore, may respond to environmental conditions with biochemical and/or metabolic changes. Such changes have been reported to include phenotypic changes, wherein a microorganism responds to environmental changes by expressing certain sets of genes, resulting in functional and structural adjustments within the microorganism to adapt to the changing environmental conditions. Microorganisms have a wide range of regulatory mechanisms to sense and respond to changing environmental conditions, and this confers on them a high level of versatility.

[0004] Replacement of traditional chemical production processes relying on, for example fossil fuels and/or potentially toxic chemicals, with environmentally friendly and/or sustainable solutions is being considered, including work to identify suitable building blocks for such use in the manufacturing of such chemicals. In Cupriavidus necator, polyhydroxybutyrate (PHB) is a key intracellular carbon and energy storage compound enabling cells to survive periods of starvation, and other stressful conditions. In response to such environmental variables, regulators alter global gene expression and metabolism and under appropriate conditions this can result in the storage of carbon and energy as PHB (Anderson and Dawes, Microbiol. Rev. 1990 54: 450-72).

SUMMARY

[0005] Methods for increasing product yield of organisms and organisms capable of increased product yield are provided.

[0006] An aspect of the present invention relates to methods for increasing carbon-based chemical product yield in an organism. The methods comprise modulating activity of one or more polypeptides or functional fragments thereof involved in a stringent response and/or in a regulatory network of an organism selected from a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto.

[0007] In one nonlimiting embodiment, one or more modified genes limiting intracellular glutamine are modified.

[0008] In one nonlimiting embodiment, one or more genes selected from glutamate dehydrogenase (gdhA; H16_A0471, H16_B1945, or H16_A1356), glutamine synthetase (glnA; H16_A2335, H16_B0618 or H16_B2191) or glutamate synthase (gltAB; H16_A3430 or H16_A3431) are modified.

[0009] In one nonlimiting embodiment, one or more genes selected from GlnB (H16_A0320 and H16_0750), NtrB (H16_A2333), NtrC (H16_A2332), PtsN (H16_A0384), GlnD (H16_A2057), spoT1 (H16_A0955) and/or spoT2 (H16_A1337) are modified.

[0010] In one nonlimiting embodiment, one or more genes selected from NtrC (H16_A2332), RpoN/sigma 54 (H16_A0387), sigma 54 modulation protein S30EA (H16_A0386), or homologs thereof are modified.

[0011] In one nonlimiting embodiment, one or more genes affecting a signaling pathway leading to increased (p)ppGpp levels and induction of the stringent response in non-limiting culture conditions are modified.

[0012] In one nonlimiting embodiment, one or more modified genes involved in the regulatory network selected from FadP (H16_A0459), PsrA (H16_A3736), LldR (H16_B0094), GlpR (H16_A2504), AccR/BphQ (H16_A1372 or H16_A1373) and/or HexR (H16_A1177 or H16_B1210) and/or are involved in a regulatory network involving glycerol, the Ntr system and/or a sucA riboswitch are modified.

[0013] Another aspect of the present invention relates to a nonnaturally occurring organism capable of yielding a carbon-based chemical product. These nonnaturally occurring organisms comprise a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto and a modification to one or more polypeptides or functional fragments thereof encoded by one or more genes involved in a stringent response and/or in a regulatory network of the organism.

[0014] In one nonlimiting embodiment, one or more genes limiting intracellular glutamine are modified in the nonnaturally occurring organism.

[0015] In one nonlimiting embodiment, one or more genes selected from glutamate dehydrogenase (gdhA; H16_A0471, H16_B1945, or H16_A1356), glutamine synthetase (glnA; H16_A2335, H16_B0618 or H16_B2191) or glutamate synthase (gltAB; H16_A3430 or H16_A3431) are modified.

[0016] In one nonlimiting embodiment, one or more genes selected from GlnB (H16_A0320 and H16_0750) NtrB (H16_A2333), NtrC (H16_A2332), PtsN (H16_A0384), GlnD (H16_A2057), spoT1 (H16_A0955) and/or spoT2 (H16_A1337) are modified.

[0017] In one nonlimiting embodiment, one or more genes selected from NtrC (H16_A2332), RpoN/sigma 54 (H16_A0387), sigma 54 modulation protein S30EA (H16_A0386), or homologs thereof are modified.

[0018] In one nonlimiting embodiment, one or more genes affecting a signaling pathway leading to increased (p)ppGpp levels and induction of the stringent response in non-limiting culture conditions are modified.

[0019] In one nonlimiting embodiment, one or more modified genes involved in the regulatory network selected from FadP (H16_A0459), PsrA (H16_A3736), LldR (H16_B0094), GlpR (H16_A2504), AccR/BphQ (H16_A1372 or H16_A1373) and/or HexR (H16_A1177 or H16_B1210) and/or are involved in a regulatory network involving glycerol, the Ntr system and/or a sucA riboswitch are modified.

[0020] Yet another aspect of the present invention relates to methods for producing a carbon-based chemical product. In these methods, a nonnaturally occurring organism of the present invention is fermented with a carbon source.

BRIEF DESCRIPTION OF THE FIGURES

[0021] FIGS. 1A through 1D are bargraphs showing accumulation of biomass (OD at 600 nm; FIG. 1A) and normalised isopropanol (IPA; FIG. 1B) after 48 hours of feeding regime. "Control" is a recombinant mutant of C. necator H16 where the PHBs pathway was replaced by the IPA pathway. .DELTA.FadP is a recombinant mutant of C. necator H16 where the PHBs pathway was replaced by the IPA pathway and a fatty acid degradation regulator, FadP, was deleted. Both strains have consumed almost all fructose introduced in the system (FIG. 1C) and have exhausted all the nitrogen available (FIG. 1D). Data represent average of 3 biological replicates and error bars represent twice the standard deviation of the datasets.

DETAILED DESCRIPTION

[0022] This disclosure provides methods for increasing carbon-based chemical product yield in an organism via modification to one or more genes involved in a stringent response or in regulatory networks.

[0023] By "modification", "modifying" or "modify" for purposes of the present invention, it is meant that the gene is deleted, mutated, overexpressed or attenuated.

[0024] In certain aspects, the organism is modified by altering, engineering, or introducing one or more nucleic acid sequences within the organism. The altering of modifying of the nucleic acid sequences can be, for example and without limitation, via genetic engineering, by adaptive mutation, or by selective isolation of naturally occurring mutant strains.

[0025] In some nonlimiting embodiments, one or more enzymes or nucleic acids of the organism are modified via non-direct or rational enzyme design approaches with aims of improving activity, improving specificity, reducing feedback inhibition, reducing repression, improving enzyme solubility, changing stereo-specificity, or changing co-factor specificity. In some embodiments, the enzymes in the pathways outlined herein can be gene dosed (i.e., overexpressed by having a plurality of copies of the gene in the host organism), into the resulting genetically modified organism via episomal or chromosomal integration approaches. In some nonlimiting embodiments, genome-scale system biology techniques such as Flux Balance Analysis can be utilized to devise genome scale attenuation or knockout strategies for directing carbon flux. Attenuation strategies include, but are not limited to, the use of transposons, homologous recombination (double cross-over approach), mutagenesis, enzyme inhibitors, and RNA interference (RNAi). In some embodiments, fluxomic, metabolomic and transcriptomal data can be utilized to inform or support genome-scale system biology techniques, thereby devising genome-scale attenuation or knockout strategies in directing carbon flux. In some embodiments, the tolerance of the host microorganism to high concentrations of the extracellular product can be improved through continuous cultivation in a selective environment.

[0026] The modified nucleic acid sequences of the organism can include, for example, one or more enzymes, one or more promoters, one or more transcription factors, or combinations thereof. The modifications can be to nucleic acids encoding polypeptides functioning as a transhydrogenase, reductase, dehydrogenase, or hydrogenase enzyme or functional fragments thereof. The modifications can be to nucleic acids not directly involved in encoding polypeptides functioning as a transhydrogenase, reductase, dehydrogenase, or hydrogenase enzyme or functional fragments thereof, but indirectly affecting the polypeptides through the interconnected metabolic network and metabolic control strategy of the organism. The modification of the nucleic acid sequences can include one or more deletions, one or more substitutions, one or more insertions, or combinations thereof.

[0027] Enzymes with substitutions will generally have not more than 50 (e.g., not more than 1, not more than 2, not more than 3, not more than 4, not more than 5, not more than 6, not more than 7, not more than 8, not more than 9, not more than 10, not more than 12, not more than 15, not more than 20, not more than 25, not more than 30, not more than 35, not more than 40, or not more than 50) amino acid substitutions (e.g., conservative or non-conservative substitutions). This applies to any of the enzymes described herein and functional fragments thereof. A conservative substitution is a substitution of one amino acid for another with similar characteristics. Conservative substitutions include substitutions within the following groups: valine, alanine and glycine; leucine, valine, and isoleucine; aspartic acid and glutamic acid; asparagine and glutamine; serine, cysteine, and threonine; lysine and arginine; and phenylalanine and tyrosine. The nonpolar hydrophobic amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine and glutamine. The positively charged (basic) amino acids include arginine, lysine and histidine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Any substitution of one member of the above-mentioned polar, basic, or acidic groups by another member of the same group can be deemed a conservative substitution. In contrast, a non-conservative substitution is a substitution of one amino acid for another with dissimilar characteristics. Deletion variants can, for example, lack 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid segments (of two or more amino acids) or non-contiguous single amino acids.

[0028] In one nonlimiting embodiment, modification of the organism is carried out by allele exchange. In this embodiment, genome edits are made in a Cupriavidus or Ralstonia organism with perturbed PHB synthesis or an organism with properties similar thereto by allele exchange (also referred to as allelic exchange). In one non-limiting embodiment, the organism is a AphaCAB H16 C. necator strain generated using allele exchange.

[0029] The term `allele` is often used interchangeably with the term `gene` more generally, and refers to a defined genomic locus. In allele exchange, a specific run of DNA sequence (i.e., the native allele) in a genome of an organism is literally exchanged for a recombinant, mutant, or synthetic run of DNA sequence (i.e., the recombinant allele). Depending on the nature of the recombinant allele, this allele exchange can result in a gene deletion, a gene substitution, or a gene insertion.

[0030] In one nonlimiting embodiment, recombinant/synthetic alleles can be constructed via gene synthesis and/or standard molecular biology techniques. These alleles are then cloned into a plasmid vector for transfer into the organism and execution of the allele exchange procedure.

[0031] In some nonlimiting embodiments, the organism is modified to include one or more exogenous nucleic acid sequences.

[0032] The term "exogenous" as used herein with reference to a nucleic acid (or a protein) and an organism refers to a nucleic acid that does not occur in (and cannot be obtained from) a cell of that particular type as it is found in nature or a protein encoded by such a nucleic acid. Thus, a non-naturally-occurring nucleic acid is considered to be exogenous to a host once in the host. It is important to note that non-naturally-occurring nucleic acids can contain nucleic acid subsequences or fragments of nucleic acid sequences that are found in nature provided the nucleic acid as a whole does not exist in nature. For example, a nucleic acid molecule containing a genomic DNA sequence within an expression vector is non-naturally-occurring nucleic acid, and thus is exogenous to a host cell once introduced into the host, since that nucleic acid molecule as a whole (genomic DNA plus vector DNA) does not exist in nature. Thus, any vector, autonomously replicating plasmid, or virus (e.g., retrovirus, adenovirus, or herpes virus) that as a whole does not exist in nature is considered to be non-naturally-occurring nucleic acid. It follows that genomic DNA fragments produced by PCR or restriction endonuclease treatment as well as cDNAs are considered to be non-naturally-occurring nucleic acid since they exist as separate molecules not found in nature. It also follows that any nucleic acid containing a promoter sequence and polypeptide-encoding sequence (e.g., cDNA or genomic DNA) in an arrangement not found in nature is non-naturally-occurring nucleic acid. A nucleic acid that is naturally-occurring can be exogenous to a particular host microorganism. For example, an entire chromosome isolated from a cell of yeast x is an exogenous nucleic acid with respect to a cell of yeast y once that chromosome is introduced into a cell of yeast y.

[0033] In contrast, the term "endogenous" as used herein with reference to a nucleic acid (e.g., a gene) (or a protein) and a host refers to a nucleic acid (or protein) that does occur in (and can be obtained from) that particular host as it is found in nature. Moreover, a cell "endogenously expressing" a nucleic acid (or protein) expresses that nucleic acid (or protein) as does a host of the same particular type as it is found in nature. Moreover, a host "endogenously producing" or that "endogenously produces" a nucleic acid, protein, or other compound produces that nucleic acid, protein, or compound as does a host of the same particular type as it is found in nature.

[0034] In certain aspects, the organism is modified to include one or more functional fragments of enzymes, other polypeptides, or nucleic acids. The phrase "functional fragment" as used herein refers to a peptide fragment of a polypeptide or a nucleic acid sequence fragment encoding a peptide fragment of a polypeptide that has at least 25%, e.g., at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or at least 100% of the activity of the corresponding mature, full-length, polypeptide. The functional fragment can generally, but not always, be comprised of a continuous region of the polypeptide, wherein the region has functional activity.

[0035] The stringent response is triggered under stressful conditions, for example, nutrient starvation, and allows the cell to survive under different extreme environmental conditions. In one nonlimiting embodiment of the present invention, one or more genes involved in a stringent response of the organism are modified.

[0036] A nonlimiting example of altering genome regulators for stringent responses is limited nitrogen.

[0037] For example, in conditions of nitrogen deficiency levels of glutamine are low resulting in the uridylation of GlnB (H16_A0750, H16_A0320) by GlnD (H16_A2057). Unmodified GlnB stimulates the phosphatase activity of the kinase/phosphatase NtrB (H16_A2333), thus a consequence of GlnB uridylation is the phosphorylation of NtrC (H16_A2332), an enhancer binding protein, to its transcriptionally active form by the kinase NtrB. Activated NtrC binds to promoter regions and together with sigma 54 (RpoN H16_A0387) activates transcription from sigma 54 dependent promoters such as the promoter for glnA (H16_A2335), and in turn various effector response genes (Sacamboio et al. Scientific Reports 2017 7:13546; Lardi et al. Appl. Environ. Microbiol. 2015 81(12): 4077-4089; Persuhn et al. FEMS Microbiology Letters 2000 192 217-221; Liu et al. PLoS One 2017 12(6):e0179037). In E. coli this set of genes includes relA, providing a direct link between nitrogen limitation and the (p)ppGpp levels (Brown et al. Nat. Comm. 2014 5:4115). In one nonlimiting embodiment of the present invention, altered glutamine levels (nitrogen signal molecule) can be achieved by modifying glutamate dehydrogenase (gdhA1, gdhA2-H16_A0471; H16_B1945), glutamine synthetase (ginA1, glnA2, glnA3-H16_A2335; H16_B0618; H16_B2191), and/or glutamate synthase (gltD, gltB1-H16_A3430; H16_A3431). In one nonlimiting embodiment, the gene H16_A2335 is mutated or deleted.

[0038] In addition, the nitrogen regulatory system is encoded by uridyltransferase (glnD H16_A2057), trimeric signaling protein PII (glnB H16_A0750, H16_A0320), as well as the kinase NtrB (H16_A2333) and enhancer binding protein NtrC (H16_A2332). Deletion of the ntrC gene resulted in excess PHB production and an increase in the NADPH/NADP+ ratio in Herbaspirillum seropedicae (Sacamboio et al. Scientific Reports 2017 7:13546), while in Azospirillum brasilense deletion of ntrC or glnD results in higher PHB production (Sun et al. Appl. Environ. Microbiol. 2002 68(2):985-988; Sun et al. Appl. Environ. Microbiol. 2000 66(1):113-117). In Paracoccus denitrificans, mutation of the NtrB sensor kinase resulted in a strain with elevated PHB and acetyl-CoA levels (Olaya-Abril et al. FEMS Microbiology Letters 2008 365:fnx251).

[0039] In one nonlimiting embodiment of the present invention, one or more genes selected from glnB (H16_A0320 and/or H16_A0750), ntrB (H16_A2333), ntrC (H16_A2332), and glnD (H16_A2057) are modified.

[0040] Nitrogen sigma factor RpoN (sigma 54) has also been implicated in the response to nitrogen limitation and PHB production (Lardi et al. Appl. Environ. Microbiol. 2015 81(12)4077-4089). In one nonlimiting embodiment of the present invention, expression of sigma 54 (H16_A0387) or another form, sigma 54 modulation protein S30EA (H16_A0386) is attenuated.

[0041] The alarmone (p)ppGpp is another important signal molecule that induces stringent response in many bacterial species. It has been shown to alter transcription and is associated with stresses such as nitrogen limitation (Juengert et al. Applied and Environmental Microbiology 2017 83(13):e00755-17; Karstens et al. Microbiology 2014 160:711-722; Hauryliuk et al. Nature Reviews Microbiology 2015 13:298-309). The alarmone ppGpp is a key signaling molecule in bacteria and is involved in regulating growth and stress responses. (p)ppGpp is synthesized from GTP by the action of RelA/Rel/SpoT enzymes which can catalyze the phosphorylation of GTP. Hydrolysis of (p)ppGpp back to GTP is catalyzed by Rel and SpoT (but not RelA). C. necator has SpoT-like and RelA-like genes. (p)ppGpp can exert its regulatory effect by several mechanisms including, but not limited to, alteration of GTP pool (changes promoter preference of RNAP), binding to RNAP; inhibition of protein biosynthesis, inhibition of DNA replication, inhibition of polyphosphate metabolism and inhibition of acid stress response. In C. necator (p)ppGpp levels are regulated by the activities of SpoT1 (p)ppGpp synthase/hydrolase (H16_A0955) and SpoT2 (p)ppGpp synthase (H16_A1337). C. necator strains lacking SpoT1 have increased levels of (p)ppGpp and increased PHB accumulation, while strains lacking both SpoT1 and SpoT2 have no detectable (p)ppGpp and low PHB levels (Juengert et al. Applied and Environmental Microbiology 2017 83(13):e00755-17).

[0042] The phosphotransferase system in C. necator is part of the system that regulates PHB production. In C. necator this system consists of EI (Ptsl H16_A0326), HPr (PtsH H16_A0325), EIIA.sup.Ntr(PtsN H16_A0384), EIIA.sup.Man(PtsM H16_A0324) and HprK (H16_A0383) (Krausse et al. J Mol Microbiol Biotechnol 2009 17:146-152; Kaddor and Steinbuchel. Appl. Environ. Microbiol. 2011 77:3582-3590). Knockout of ptsN in C. necator results in increased accumulation of PHB, while knockouts of ptsI and ptsH result in a decrease in PHB. Unphosphorylated PtsN has also been shown to interact with the ppGpp synthase/hydrolase SpoT1 thus influencing (p)ppGpp levels and the stringent response (Karstens et al. Microbiology 2014 160:711-722).

[0043] In one nonlimiting embodiment of the present invention, the organism is modified to express either or both of the natural and/or non-native spoT genes (H16_A0955 and H16_A1337). In another nonlimiting embodiment of the present invention, spoT1 (H16_A0955) and/or spoT2 (H16_A1337) are mutated or deleted.

[0044] In one nonlimiting embodiment of the present invention, the organism is modified to attenuate expression of ptsN (H16_A0384).

[0045] In one nonlimiting embodiment of the present invention, one or more genes involved in a regulatory network of the organism are modified.

[0046] In one nonlimiting embodiment, FadP (H16_A0459) is modified. FadP is a TetR-like regulator located in the fatty acid degradation operon in C. necator H16 (H16_A0459-465) and is a potential repressor of fatty acid degradation pathways (Kazakov et al. Journal of Bacteriology 2009 191:52-64). Deletion of H16_A0459 is expected to upregulate .beta.-oxidation pathways which is desired for improved growth and/or utilization of organic acids and mixtures thereof.

[0047] Overexpression of H16_A0459 is expected to downregulate .beta.-oxidation pathways, which is desired for production of fatty acid derived products. The predicted promoter motif of FadP is AATNGWACGAYCGTKCKNWT (SEQ ID NO:1). FadP is expected to regulate for example, but not limited to, H16_A0217 (thioesterase), H16_A0459-H160464 (beta-oxidation operon), H16_A0814-16 (electron transfer flavoprotein beta, electron transfer flavoprotein alpha, acyl-CoA dehydrogenase), H16_A1066-1068 (2.times.acyl-CoA dehydrogenase), H16_A1102-3 (3-HO-acyl-CoA dehydrogenase, enoyl-CoA hydratase), H16_A1445 (BktB), H16_A1519 (acyl-CoA ligase), H16_A1526-31 (beta-oxidation operon), H16_A2794 (acyl-CoA ligase) and H16_A3288 (acyl-CoA ligase).

[0048] Organisms with a mutation in the fadP gene are expected to exhibit improved utilization of fatty acids/mixtures containing fatty acids/mixotrophic growth. Further, removal of a FadP binding site in promoter regions of genes/operons will deregulate specific pathways or groups of genes thereby improving utilization of fatty acids/mixtures containing fatty acids/mixotrophic growth. Organisms with increased FadP expression and decreased beta-oxidation are expected to exhibit reduced degradation of fatty acids thus improving production of fatty acids/fatty acid derived products. Organisms with increased FadP expression and decreased beta-oxidation are also expected to reduce degradation of fatty acids for cleanup of mixed waste stream (e.g. removal of non-diacids from NVR to leave just adipic and/or glutaric acids). Organisms with increased FadP expression with mutated FadP binding sites to simultaneously reduce expression of specific beta-oxidation pathway genes and increase expression of others are useful in, for example, but not limited to, clean up of mixed waste stream (as above) or for production of fatty acid derived molecules while using, for example, but not limited to, oils or NVR as growth substrate.

[0049] In one nonlimiting embodiment, PsrA (H16_A3736) is modified. PsrA is a TetR family regulator and also regulates fatty acid pathways. PsrA is expected to regulate the fatty acid biosynthesis (FAB) operon (H16_A2569-2565) and its deletion is expected to upregulate FAB pathways resulting in more fatty acid production. Alternatively, when acting as a repressor, overexpression could reduce the FAB pathway.

[0050] Organisms with mutation in PsrA may exhibit increased production of fatty acids and fatty acid derived molecules or decreased production of fatty acids with increased production of other desired products via redirection of flux and/or increased availability of acetyl-CoA for other pathways.

[0051] In one nonlimiting embodiment, LldR is modified. LldR (H16_B0094) is a GntR family regulator and is a potential repressor of lactate dehydrogenase expression (H16_B0460, H16_B1817). H16_B0094 is adjacent to an operon which includes L-lactate permease (H16_B0090). Deletion of LldR is expected to upregulate lactate catabolism (Gao et al. Journal of Bacteriology 2012 194:2687-269; Leyn et al. Microbial Genomics 2016 1-15).

[0052] Organisms with mutated LldR or LldR binding sites are expected to exhibit increased utilization of lactate and production of products comprising organic acid mixtures that include lactate. Organisms with increased expression of LldR are expected to exhibit decreased utilization of lactate for improved production of lactate and lactate derived products.

[0053] In one nonlimiting embodiment, one or more genes involved in the regulatory network involving glycerol are modified. In one embodiment, the gene is GlpR (H16_A2504), which is co-localized with genes for glycerol kinase (glpK H16_A2507), glycerol-3-phosphate dehydrogenase (glpD H16_A2508), and an ABC transporter system (H16_A2498-2503) that is predicted to transport glycerol. The ABC transporter system encoded by H16_A2498-2503 has high identity (49%-73%) to a characterized glycerol transport system from Rhizobium leguminosarum (Ding et al. Microbiology 2012 158:1369-1378). In one nonlimiting embodiment, GlpR is deleted or expression is reduced thereby increasing glycerol utilization and/or improving utilization of mixtures containing glycerol. In one nonlimiting embodiment, GlpR is overexpressed thereby decreasing glycerol utilization. Modification of one or more genes involved in the regulatory network involving glycerol is expected to improve production of glycerol derived products such as 3-hydroxypropionate and 1,3,-propanediol. Further, organisms with glycerol uptake but no glycerol metabolism can be produced.

[0054] In one nonlimiting embodiment, one or more genes involved in the regulatory network involving catabolism of aromatic compounds are modified. In one nonlimiting embodiment, AccR and/or its corresponding sensor kinase is modified (Valderrama et al. Journal of Biological Chemistry 2014 289(4):1892-1904). In one nonlimiting embodiment, H16_A1372, the AccR response regulator and/or H16_A1373, its corresponding sensor kinase, are modified. Phosphorylation of AccR causes repression of genes involved in aromatic hydrocarbon catabolism and its modification is predicted to affect aromatic hydrocarbon degradation pathways. Further, AccR is predicted to regulate promoters for, for example, but not limited to, succinate dehydrogenase (H16_A2629-32), benzoate regulatory genes (H16_A1411; H16_B1915) and benzoyl-coA ligase (H16_A1412).

[0055] Organisms with mutated AccR and mutations in AccR binding sites are expected to exhibit improved biomass formation and improved utilization and formation of products of aromatic hydrocarbons and mixtures containing aromatic hydrocarbons. A nonlimiting example is terephthalic acid. Organisms with increased AccR, increased kinase and/or AccR with a super repressor mutation are expected to exhibit decreased utilization of aromatics and improved production of aromatic derived molecules.

[0056] Mutation of Asp.sup.60 to the phosphomimetic Glu in the Azoarcus AccR results in a constitutively active AccR that is a super repressor of target genes (Valderrama et al. Journal of Biological Chemistry 2014 289(4):1892-1904). Organisms of the present invention modified to reduce or delete AccR are expected to exhibit increased aromatic utilization and mixtures containing aromatics. Organisms of the present invention with increased AccR expression or with a super repressor AccR (e.g. Asp.sup.63 to Glu in H16_A1372) are expected to exhibit decreased aromatic utilization.

[0057] In one nonlimiting embodiment, HexR (Leyn et al. Microbial Genomics 2016 1-15; Leyn et al. Journal of Biological Chemistry 2011 286(41): 35782-35794) is modified. In one nonlimiting embodiment, HexR genes from C. necator H16, H16_A1177 and/or H16_B1210 are modified. H16_A1177 is divergently transcribed from genes encoding the ED pathway enzymes phosphogluconate dehydratase (edd H16_A1178) and gluconate kinase (H16_A1179). H16_B1210 is divergently transcribed from genes encoding the ED pathway enzymes 2-keto-3-deoxygluconate kinase (H16_B1212) and 2-keto-3-deoxy-6-phosphogluconate aldolase (eda H16_B1213). HexR is an Entner-Doudoroff (ED) pathway regulator, the major sugar degradation route in C. necator. Overexpression of HexR is expected to alter carbon preferences (gluconate/glucose) in organisms improving or reducing flux through the ED pathway.

[0058] In one nonlimiting embodiment, one or more genes involved in the regulatory network involving riboswitches are modified. Riboswitches are described, for example, by Weinberg et al. (Nucleic Acids Research 2007 35:4809-4819). In one nonlimiting embodiment, a sucA riboswitch is modified. In one nonlimiting embodiment, a riboswitch comprising the nucleic acid sequence of TTGTTTGCGATCCGCTAACCGGTCAAGCCGTGTCGCGGAAGGTTGAATAACCCGCTGAACTC CGGCATTCCCGGAGAATAGTGAGCGTCCCATGATG (SEQ ID NO:2) is modified. In one nonlimiting embodiment, a riboswitch upstream of H16_A2325, 2-oxoglutarate dehydrogenase, is modified.

[0059] Organism with mutations of a sucA riboswitch and deregulation of 2-oxo-glutarate dehydrogenase are expected to exhibit increased production of products derived from TCA cycle intermediates.

[0060] Nonlimiting examples of nucleic acid and amino acid sequences for the above-described genes are set forth in the Sequence Appendix. However, as will be understood by the skilled artisan upon reading this disclosure, the present invention is not limited to these particular gene sequences and is also inclusive of nucleic acid and amino acid sequences for polypeptides with similar enzymatic activities exhibiting at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% sequence identity to sequences set forth in the Sequence Appendix and functional fragments thereof.

[0061] The percent identity (and homology) between two amino acid sequences as disclosed herein can be determined as follows. First, the amino acid sequences are aligned using the BLAST 2 Sequences (B12seq) program from the stand-alone version of BLAST containing Blast+2.9.0. This stand-alone version of BLAST can be obtained from the U.S. government's National Center for Biotechnology Information web site (www with the extension ncbi.nlm.nih.gov). Instructions explaining how to use the B12seq program can be found in the readme file accompanying BLASTZ. B12seq performs a comparison between two amino acid sequences using the BLASTP algorithm. To compare two amino acid sequences, the options of B12seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seq1.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\B12seq c:\seq1.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two compared sequences share homology (identity), then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology (identity), then the designated output file will not present aligned sequences. Similar procedures can be followed for nucleic acid sequences except that blastn is used.

[0062] Once aligned, the number of matches is determined by counting the number of positions where an identical amino acid residue is presented in both sequences. The percent identity (and homology) is determined by dividing the number of matches by the length of the full-length polypeptide amino acid sequence followed by multiplying the resulting value by 100. It is noted that the percent identity (homology) value is rounded to the nearest tenth. For example, 90.11, 90.12, 90.13, and 90.14 is rounded down to 90.1, while 90.15, 90.16, 90.17, 90.18, and 90.19 is rounded up to 90.2. It also is noted that the length value will always be an integer. It will be appreciated that a number of nucleic acids can encode a polypeptide having a particular amino acid sequence. The degeneracy of the genetic code is well known to the art; i.e., for many amino acids, there is more than one nucleotide triplet that serves as the codon for the amino acid. For example, codons in the coding sequence for a given enzyme can be modified such that optimal expression in a particular species (e.g., bacteria or fungus) is obtained, using appropriate codon bias tables for that species.

[0063] By "carbon-based chemical product" as used herein, it is meant to include C3 to C12 alkenes, alcohols, diols, monoacids, diacids, hydroxyacids, amino acids and diamines. In one nonlimiting embodiment, the carbon-based chemical product may be any C6-C12 difunctional aliphatic fatty acid or derivative thereof including, but not limited to, C6-C12 amino acids, C6-C12 diamines, C6-C12 hydroxyacids, C6-C12 diols, and C6-C12 diacids. Nonlimiting examples of carbon-based chemical products produced in accordance with this disclosure include 1,3-propanediol, 1,2-propanediol, methionine, threonine, lysine, glutamic acid, tryptophan, aspartic acid, leucine, isoleucine, valine, citric acid, maleic acid, succinic acid, isoprene, linalool, limonene, 3-hydroxypropanoic acid, malonic acid, lactic acid, n-butanol, 2-butanone, butadiene, 2-3 butanediol, 1-3 butanediol, benzoic acid, 1,4-benzenediamine, benzeneamine, pyridine, vanillin, hydroquinone, 1,4-diaminobutane, 2-hydroxyisobutyric acid, itaconic acid, 3-hydroxybutyrate and nylon intermediates.

[0064] In some nonlimiting embodiments, the organism has been modified to exhibit an increased synthesis of the extracellular product relative to that of the corresponding wild type organism.

[0065] In some nonlimiting embodiments, the carbon-based chemical product includes pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, 1,7-heptanediol, or a combination thereof. Additional descriptions of the synthesis of these carbon-based chemical products with Ralstonia, Cupriavidus, or an organism similar thereto can be found in U.S. Pat. No. 10,196,657, the disclosure of which is incorporated by reference herein in its entirety for all purposes.

[0066] In some nonlimiting embodiments, the carbon-based chemical product includes 1,4-butanediol, putrescine, 4-hydroxybutyrate, 4-aminobutyrate, or a combination thereof. Additional descriptions of the synthesis of these carbon-based chemical products with Ralstonia, Cupriavidus, or an organism related thereto can be found in U.S. Pat. Nos. 10,072,150 and 9,637,764, the disclosures of which are incorporated by reference herein in their entirety for all purposes.

[0067] In some nonlimiting embodiments, the carbon-based chemical product includes glutaric acid, 5-aminopentanoic acid, cadaverine (also known as 1,5 pentanediamine), 5-hydroxypentanoic acid, 1,5-pentanediol, glutarate semialdehyde (also known as 5-oxopentanoate), or a combination thereof. Additional descriptions of the synthesis of these carbon-based chemical products with Ralstonia, Cupriavidus, or an organism related thereto can be found in U.S. Pat. No. 9,920,339, the disclosure of which is incorporated by reference herein in its entirety for all purposes.

[0068] In some nonlimiting embodiments, the carbon-based chemical product includes isoprene. Additional descriptions of the synthesis of this carbon-based chemical product with Ralstonia, Cupriavidus, or an organism related thereto can be found in U.S. Pat. No. 9,862,973, the disclosure of which is incorporated by reference herein in its entirety for all purposes.

[0069] In some nonlimiting embodiments, the carbon-based chemical product includes adipic acid, 6-aminohexanoic acid, hexamethylenediamine, caprolactam, 1,6-hexanediol, or a combination thereof. Additional descriptions of the synthesis of these carbon-based chemical products with Ralstonia, Cupriavidus, or an organism related thereto can be found in U.S. Pat. No. 9,580,733, the disclosure of which is incorporated by reference herein in its entirety for all purposes.

[0070] For products of the present invention containing carboxylic acid groups such as organic monoacids, hydroxyacids, aminoacids and dicarboxylic acids, these products may be formed or converted to their ionic salt form when an acidic proton present in the parent product either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and/or bicarbonate, sodium hydroxide, ammonia and the like. The salt can be isolated as is from the system as the salt or converted to the free acid by reducing the pH to below the lowest pKa through addition of acid or treatment with an acidic ion exchange resin.

[0071] For products of the present invention containing amine groups such as but not limited to organic amines, amino acids and diamine, these products may be formed or converted to their ionic salt form by addition of an acidic proton to the amine to form the ammonium salt, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid or muconic acid. The salt can be isolated as is from the system as a salt or converted to the free amine by raising the pH to above the lowest pKa through addition of base or treatment with a basic ion exchange resin. Acceptable inorganic bases are known in the art and include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and/or bicarbonate, ammonia, sodium hydroxide, and the like.

[0072] For products of the present invention containing both amine groups and carboxylic acid groups such as but not limited to amino acids, these products may be formed or converted to their ionic salt form by either 1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid or muconic acid. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like or 2) when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base. Acceptable organic bases are known in the art and include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. Acceptable inorganic bases are known in the art and include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and/or bicarbonate, ammonia, sodium hydroxide, and the like. The salt can be isolated as is from the system or converted to the free acid by reducing the pH to below the lowest pKa through addition of acid or treatment with an acidic ion exchange resin.

[0073] Nonnaturally occurring organism produced and used in accordance with the present invention are selected from a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto.

[0074] For purposes of the present invention, by "diminishing" or "diminished" polyhydroxybutyrate synthesis, it is meant that the organism is altered to synthesize less polyhydroxybutyrate as compared to an unaltered wild-type organism of the same species. Organisms used in this disclosure can exhibit at least 20%, 25%, 30%, 40%, 50% or even greater decreased polyhydroxybutyrate synthesis as compared to an unperturbed wild-type organism of the same species.

[0075] Nonlimiting examples of species of Cupriavidus or Ralstonia useful in accordance with this disclosure include Cupriavidus necator, Cupriavidus metallidurans, Cupriavidus taiwanensis, Cupriavidus pinatubonensis, Cupriavidus basilensis and Ralstonia pickettii.

[0076] C. necator (also referred to as Hydrogenomonas eutrophus, Alcaligenes eutropha, Ralstonia eutropha, and Wautersia eutropha) is a Gram-negative, flagellated soil bacterium of the Betaproteobacteria class. This hydrogen-oxidizing bacterium is capable of growing at the interface of anaerobic and aerobic environments and easily adapts between heterotrophic and autotrophic lifestyles. Sources of energy for the bacterium include both organic compounds and hydrogen. Additional properties of C. necator include microaerophilicity, copper resistance (Makar, N. S. & Casida, L. E. Int. J. of Systematic Bacteriology 1987 37(4): 323-326), bacterial predation (Byrd et al. Can J Microbiol 1985 31:1157-1163; Sillman, C. E. & Casida, L. E. Can J Microbial 1986 32:760-762; Zeph, L. E. & Casida, L. E. Applied and Environmental Microbiology 1986 52(4):819-823) and polyhydroxybutyrate (PHB) synthesis. In addition, the cells have been reported to be capable of either aerobic or nitrate dependent anaerobic growth. A nonlimiting example of a C. necator organism useful in the present invention is a C. necator of the H16 strain. In one nonlimiting embodiment, a C. necator host of the H16 strain with at least a portion of the phaC1AB1 gene locus knocked out (AphaCAB) is used. In one nonlimiting embodiment, the organism is further modified to eliminate phaCAB, involved in PHBs production and/or H16-A0006-9 encoding endonucleases thereby improving transformation efficiency as described in U.S. patent application Ser. No. 15/717,216, teachings of which are incorporated herein by reference. However, other means of eliminating PHB synthesis are included within the scope of the invention.

[0077] By "an organism with properties similar thereto" it is meant an organism having one or more of the above-mentioned properties of C. necator.

[0078] In the process described herein, a fermentation strategy can be used that entails anaerobic, micro-aerobic or aerobic cultivation coupled with nutrient limitation such as iron, sulphate, nitrogen, potassium, oxygen, phosphorus, carbon and/or or NADP limitations, gradients thereof and any combinations thereof.

[0079] A cell retention strategy using a ceramic hollow fiber membrane can also be employed to achieve and maintain a high cell density during fermentation.

[0080] The principal carbon source fed to the fermentation can derive from a biological or non-biological feedstock. In one nonlimiting embodiment, the feedstock is fed to the fermentation as a gaseous or liquid stream.

[0081] Accordingly, feedstocks for fermentation may be gases such as carbon dioxide or hydrogen; sugars such as glucose, xylose or fructose; sugar acids such as gluconate; fatty acids or fats/oils, carboxylic acids such as propionic acid, lactic acid, and formic acid; amino acids, aromatics such as phenol and benzoic acid and/or alcohols such as glycerol.

[0082] The feedstocks may be carbon sources derived from by-product or waste streams such as brewing, dairy, plant oil, ethanol, corn, soy, fish, or sugar industries or any other food or agricultural waste such as used cooking oil.

[0083] The biological feedstock can be, or can derive from, monosaccharides, disaccharides, lignocellulose, hemicellulose, cellulose, paper-pulp waste, black liquor, lignin, levulinic acid and formic acid, triglycerides, glycerol, fatty acids, agricultural waste, thin stillage, condensed distillers' solubles or waste streams from the food processing or dairy industries municipal waste such as fruit peel/pulp or whey.

[0084] The non-biological feedstock can be, or can derive from, natural gas, syngas, CO.sub.2/H.sub.2, CO, H.sub.2, O.sub.2, methanol, ethanol, waste streams from processes to produce monomers for the Nylon-66 and Nylon-6 industries such as but not limited to non-volatile residues (NVRs) and caustic wash waste streams from the cyclohexane oxidation process used to manufacture adipic acid or caprolactam or waste stream from other chemical industry processes such as, but not limited to a carbon black industry or a hydrogen-refining industry, or petrochemical industry, a nonlimiting example being a PTA-waste stream.

[0085] In one nonlimiting embodiment, at least one of the enzymatic conversions of the production method comprises gas fermentation within the modulated Ralstonia or Cupriavidus organism or other organism with properties similar thereto. In this embodiment, the gas fermentation may comprise at least one of natural gas, syngas, CO, H.sub.2, O.sub.2, CO.sub.2/H.sub.2, methanol, ethanol, non-volatile residue, caustic wash from cyclohexane oxidation processes, or waste stream from a chemical industry such as, but not limited to a carbon black industry or a hydrogen-refining industry, or petrochemical industry. In one nonlimiting embodiment, the gas fermentation comprises CO.sub.2/H.sub.2.

[0086] The methods of the present invention may further comprise recovering produced product from the organism. Once produced, any method can be used to isolate these products or derivatives or compounds related thereto. The isolation of at least one product can involve any one or more downstream processes generally known to be suitable for the at least partial separation and/or isolation of material from a reaction or bioprocess. The collection can, for example, involve centrifugations, cell disruptions, concentrations, precipitations, extractions, filtrations, crystallizations, distillations, chemical conversions, or combinations thereof. One or more biosynthetic products can be collected from the liquid or solid phase of the culture, or from the gas phase present in the headspace of a bioreactor or the off-gas.

[0087] The present invention also provides nonnaturally occurring organisms and methods for producing the nonnaturally occurring organisms modified to increase carbon-based product yield via modification to one or more genes involved in a stringent response or in regulatory networks. These nonnaturally occurring organisms exhibit increased product yield as compared to product yield in the same organism without modification to one or more genes involved in a stringent response or in regulatory networks. The nonnaturally occurring organisms are selected from a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto.

[0088] Nonlimiting examples of species of Cupriavidus or Ralstonia useful in accordance with this disclosure include Cupriavidus necator, Cupriavidus metallidurans, Cupriavidus taiwanensis, Cupriavidus pinatubonensis, Cupriavidus basilensis and Ralstonia pickettii.

[0089] In one nonlimiting embodiment, the present invention relates to a substantially pure culture of the nonnaturally occurring organism modified to comprise one or more promoters which are inducible under one or more specific limitation conditions.

[0090] As used herein, a "substantially pure culture" of an altered organism is a culture of that microorganism in which less than about 40% (i.e., less than about 35%; 30%; 25%; 20%; 15%; 10%; 5%; 2%; 1%; 0.5%; 0.25%; 0.1%; 0.01%; 0.001%; 0.0001%; or even less) of the total number of viable cells in the culture are viable cells other than the altered microorganism, e.g., bacterial, fungal (including yeast), mycoplasmal, or protozoan cells. The term "about" in this context means that the relevant percentage can be 15% of the specified percentage above or below the specified percentage. Thus, for example, about 20% can be 17% to 23%. Such a culture of nonnaturally occurring microorganisms includes the cells and a growth, storage, or transport medium. Media can be liquid, semi-solid (e.g., gelatinous media), or frozen. The culture includes the cells growing in the liquid or in/on the semi-solid medium or being stored or transported in a storage or transport medium, including a frozen storage or transport medium. The cultures are in a culture vessel or storage vessel or substrate (e.g., a culture dish, flask, or tube or a storage vial or tube).

[0091] In addition, the present invention provides bio-derived, bio-based, or fermentation-derived products produced using the methods and/or nonnaturally occurring organisms disclosed herein. Examples of such products include, but are not limited to, compositions comprising at least one bio-derived, bio-based, or fermentation-derived compound or any combination thereof, as well as molded substances, formulations and semi-solid or non-semi-solid streams comprising one or more of the bio-derived, bio-based, or fermentation-derived compounds or compositions, combinations or products thereof.

[0092] While the invention has been described in detail, in some instances making reference to a specific aspect thereof, it is apparent to one of skill in the art that various changes and modifications can be made thereto without departing from its spirit and scope. The following section provides further illustration of the methods and materials of the present invention. These Examples are illustrative only and are not intended to limit the scope of the invention in any way.

Example

[0093] Deletion of FadP Gene Increases Biomass and Isopropanol Production Under Nitrogen Limitation in Modified C. necator H16

[0094] C. necator H16 was genetically modified by deleting the polyhydroxybutyrate (PHB) pathway and the fatty acid degradation regulator, FadP (a transcriptional regulator belonging to the TetR family and is a potential repressor of fatty acid degradation pathways such as the .beta.-oxidation), and by introducing the isopropanol (IPA) pathway. The strain was grown on a fructose-based media with ammonium sulphate as a source of nitrogen. A fed batch culture was used to compare the accumulation of biomass and the production of IPA under nitrogen limiting conditions. During batch phase growth was established at 30.degree. C. and pH 6.6 in a fixed volume of 8 mL for 43 hours. During fed batch phase 5 mL of the same media lacking ammonium sulphate was fed into the cultures at a rate of 1.736 .mu.L/min. The system was sampled just before the feeding started and 48 hours later.

[0095] For each sample event, biomass (OD.sub.600) (FIG. 1A), isopropanol (FIG. 1B), residual nitrogen (FIG. 1D) and carbon FIG. 10) were determined.

[0096] These data show that at the end of the feeding regime there is more biomass present in the .DELTA.FadP strain when compared with a similar strain where the FadP gene is still present. Moreover, .DELTA.FadP strain produces higher levels of isopropanol. Both strains have consumed almost all fructose introduced in the system and have exhausted all the nitrogen available. When taken together these results are indicative of the modified strain funneling at least a portion of the potential additional carbon available towards cell growth and production of isopropanol.

TABLE-US-00001 TABLE 1 C. necator sequences Nucleic acid Amino Acid Sequence Name SEQ ID NO: SEQ ID NO: H16_A0320, glnB-1, nitrogen regulatory 3 4 protein P-II 1 H16_A0324, ptsM, PTS fructose 5 6 transporter subunit IIA H16_A0325, ptsH, phosphocarrier 7 8 protein HPr H16_A0326, ptsI, ptsI - 9 10 phosphoenolpyruvate-protein phosphotransferase H16_A0383, Hpr kinase/phosphorylase 11 12 H16_A0384, ptsN, ptsIIA - Ntr - PTS 13 14 IIA-like nitrogen-regulatory protein PtsN H16_A0386, sigma 54 modulation protein 15 16 S30EA, ribosome hibernation promoting factor H16_A0387, rpoN, sigma 54 17 18 H16_A0459, FadP, TetR - TetR/AcrR 19 20 family transcriptional regulator H16_A0471, gdhA1, Glu/Leu/Phe/Val 21 22 dehydrogenase H16_A0750, glnB-2, P-II family 23 24 nitrogen regulator H16_A0955, SpoT1, bifunctional 25 26 (p)ppGpp synthetase/guanosine-3',5'- bis(diphosphate) 3'-pyrophosphohydrolase H16_A1177, HexR-1, MurR/RpiR family 27 28 transcriptional regulator H16_A1337, SpoT2, bifunctional 29 30 (p)ppGpp synthetase/guanosine-3',5'- bis(diphosphate) 3'-pyrophosphohydrolase H16_A1356, gudB, NAD-glutamate 31 32 dehydrogenase H16_A1372, AccR, DNA-binding response 33 34 regulator H16_A1373, PAS domain-containing 35 36 sensor histidine kinase H16_A2057, glnD, [protein-PII] 37 38 uridylyltransferase H16_A2332, ntrC, Fis - nitrogen 39 40 regulation protein NR(I) H16_A2333, ntrB, PAS domain-containing 41 42 protein H16_A2335, glnA, type I glutamate- 43 44 ammonia ligase H16_A2504, GlpR, DeoR/GlpR 45 46 transcriptional regulator H16_A3430, gltD, glutamate synthase 47 48 subunit beta H16_A3431, gltB1, glutamate synthase 49 50 subunit alpha H16_A3736, PsrA, TetR/AcrR family 51 52 transcriptional regulator H16_B0094, LldR, FCD domain-containing 53 54 protein H16_B0618, glnA2, type I glutamate- 55 56 ammonia ligase H16_B1210, hexR-2, MurR/RpiR family 57 58 transcriptional regulator H16_B1945, gdhA2, NADP-specific 59 60 glutamate dehydrogenase H16_B2191, glnA3, type III glutamate- 61 62 ammonia ligase sucA, riboswitch linked to H16_A2325 2

Sequence CWU 1

1

62120DNAC. necatormisc_feature(4)..(4)n is a, c, g, or tmisc_feature(18)..(18)n is a, c, g, or t 1aatngwacga ycgtkcknwt 20297DNAC. necator 2ttgtttgcga tccgctaacc ggtcaagccg tgtcgcggaa ggttgaataa cccgctgaac 60tccggcattc ccggagaata gtgagcgtcc catgatg 973339DNAC. necator 3atgaaactca tcatcgcagt catcaagccg ttcaagctcg acgaagtgcg cgaagcgctg 60tcggacgtgg gcgtgtccgg cattaccgtg accgaagtga aaggcttcgg ccgccagaag 120ggccacaccg agctgtaccg cggcgccgaa tacatcgtcg acttcctgcc caaggtgaag 180atcgaggtgg cggtgcccga cgacgtggtc gagcgcgcca tcgaggcggt cgagaaatcg 240gcccgcaccg gcaagatcgg cgacggcaag atcttcgtgg caccgatcga gcaggtcatc 300cgcatccgca ccggcgagac cggcggcgat gccctgtga 3394112PRTC. necator 4Met Lys Leu Ile Ile Ala Val Ile Lys Pro Phe Lys Leu Asp Glu Val1 5 10 15Arg Glu Ala Leu Ser Asp Val Gly Val Ser Gly Ile Thr Val Thr Glu 20 25 30Val Lys Gly Phe Gly Arg Gln Lys Gly His Thr Glu Leu Tyr Arg Gly 35 40 45Ala Glu Tyr Ile Val Asp Phe Leu Pro Lys Val Lys Ile Glu Val Ala 50 55 60Val Pro Asp Asp Val Val Glu Arg Ala Ile Glu Ala Val Glu Lys Ser65 70 75 80Ala Arg Thr Gly Lys Ile Gly Asp Gly Lys Ile Phe Val Ala Pro Ile 85 90 95Glu Gln Val Ile Arg Ile Arg Thr Gly Glu Thr Gly Gly Asp Ala Leu 100 105 1105456DNAC. necator 5atggcaggca ttctgatcat cgcgcacacc ccgctggctt cggcgctgcg cgattgcgcc 60gcccacgtct actgcggcca gccgcagcgg ctggaatcca tcgacgtcct tcccgatgcc 120gaccccgccg tcgtgctggc cgaggccagg cgccggctgg cggccatctg cgaggacaac 180ggcgcgctgg tcctcaccga tatcttcggc gccacccccg ccaatattgc cgcacgcctg 240gccgagccgg gccgcgtgcg ggtgctggcc ggcgtcaacc ttcccatgct cgtgcgcgcg 300atctgctacc gcggcgaaaa gctcgaccag cttgccacca aggccctggc cggcggctcg 360cagggtgtgc tgcaggtcgg caccacgact gtccagaacc aaaccgcaaa ccatcccgac 420aaatatgctg cagagggaca ccaccatcat caataa 4566151PRTC. necator 6Met Ala Gly Ile Leu Ile Ile Ala His Thr Pro Leu Ala Ser Ala Leu1 5 10 15Arg Asp Cys Ala Ala His Val Tyr Cys Gly Gln Pro Gln Arg Leu Glu 20 25 30Ser Ile Asp Val Leu Pro Asp Ala Asp Pro Ala Val Val Leu Ala Glu 35 40 45Ala Arg Arg Arg Leu Ala Ala Ile Cys Glu Asp Asn Gly Ala Leu Val 50 55 60Leu Thr Asp Ile Phe Gly Ala Thr Pro Ala Asn Ile Ala Ala Arg Leu65 70 75 80Ala Glu Pro Gly Arg Val Arg Val Leu Ala Gly Val Asn Leu Pro Met 85 90 95Leu Val Arg Ala Ile Cys Tyr Arg Gly Glu Lys Leu Asp Gln Leu Ala 100 105 110Thr Lys Ala Leu Ala Gly Gly Ser Gln Gly Val Leu Gln Val Gly Thr 115 120 125Thr Thr Val Gln Asn Gln Thr Ala Asn His Pro Asp Lys Tyr Ala Ala 130 135 140Glu Gly His His His His Gln145 1507270DNAC. necator 7atgctgcaga gggacaccac catcatcaat aaactcggcc tgcatgcgcg cgcgtccgcc 60aagctgaccc agctcgccgg caactttgtc agccaggtca agatgtcccg caatggtcgc 120caggtcgacg ccaagagcat catgggcgtc atgatgctgg ccgccgggat cggctcgacg 180gtgaccctcg agaccgacgg ccccgacgag caggaggcga tggacgcgct gctggcgctg 240atcgccaacc gctttggtga gggagagtga 270889PRTC. necator 8Met Leu Gln Arg Asp Thr Thr Ile Ile Asn Lys Leu Gly Leu His Ala1 5 10 15Arg Ala Ser Ala Lys Leu Thr Gln Leu Ala Gly Asn Phe Val Ser Gln 20 25 30Val Lys Met Ser Arg Asn Gly Arg Gln Val Asp Ala Lys Ser Ile Met 35 40 45Gly Val Met Met Leu Ala Ala Gly Ile Gly Ser Thr Val Thr Leu Glu 50 55 60Thr Asp Gly Pro Asp Glu Gln Glu Ala Met Asp Ala Leu Leu Ala Leu65 70 75 80Ile Ala Asn Arg Phe Gly Glu Gly Glu 8591761DNAC. necator 9atgcctttcg ccctgcacgg catcccggtc tcgcgcggcg tcgccatcgg gcgcgcgcac 60ctgcttgcgc ccgcggcgct ggacgtgtcg cactacctgg tcgatgaaga ccagctcgac 120gccgaggtcg aacggctgcg cgcagcgcgc gccgcggtgc gggcggagct ggccgcgctc 180aagcgcgacc tgccgcgcga tgcgcccgag gagctgggcg cattcctgga cgtgcacgcg 240atgatcctcg acgacgaggc gctggcgcgc gagcccgagg ccctgatccg gggccgccgc 300tacaacgcag agtgggcact taccacgcgc ctcgaagagc tgatgcgcca gttcgacgag 360atcgaggatg aatacctgcg cgagcgcaag accgatatcc ggcaggtggt cgagcgcatc 420ctgaaggcgc tcgccggggc tccggtgctg gtgcccgcgc cggtgccggc gctggccgcc 480gacggcgagg ccgcgaccgg ggtgatcgtg gtggcccacg atatcgcccc ggccgacatg 540ctgcagttcc gccataccgt cttccacggc ttcgtcaccg acatgggcgg acgcacctcg 600cataccgcca tcgtcgcgcg cagcctggac atcccggccg cggtcggcgt gcagagcgcg 660agcgagctga tccgccagga cgactggatc atcatcgacg gcgatgccgg gctggtgatc 720gtcgacccga ccgccatcat cctggaagag taccgccacc ggcagagcga gcgcgcgctg 780gaaaagaagc gcctgcagcg gctgcggcat accccggcgg tgacgctgga cgggctggaa 840atcgacctgc tggccaatat cgagatggcc gaggacgccg gcgcggcgct ggcggccggc 900gcggtcggcg tgggcctgtt ccgttccgaa ttcctgttca tgaaccggcg cgacgagttg 960ccgggcgagg acgagcagtt ccaggcctac cgcggcgcgg tcgatgccat gcacgggctg 1020ccggtgacta tccgcaccat cgacatcggc gccgacaagc cgctcgatgc ccgcggcgat 1080gaattcgaga ccgcgctgaa cccggcgctg ggcctgcgcg cgatccgctg gtcgctgtcc 1140gagccgggca tgttcctgac ccagctgcgc gcgctgctgc gggcttcggc cttcggcccg 1200gtgcggctgc tggtgccgat gctggcgcat gccagcgaga tcgaccagac cctggcgctg 1260atcgccaagg ccaagcgcca gctcgacgag cgcggcgagg cctatgaccc gggcatgaag 1320gtcggcgcca tgatcgagat cccggcggcg gtgctgctgc tgccgctgtt cctgcgcaag 1380atggacttcc tgtccatcgg caccaacgac ctgatccagt acacgctggc catcgatcgc 1440gccgacaacg cggtggcgca cctgttcgac ccgctgcacc cggcggtgct gcagctggtg 1500gcgcgcacca tccgcgaggc caaccgcgcc ggcgtgccgg tggccgtgtg cggcgaaatg 1560gcgggcgacc cgtccatgac ccggctgctg ctgggcatgg ggctgcgcga gttctcgatg 1620cacccggcgc agctgctgcg ggtcaagcag gagatcctgc atgcccactg cgaacggctc 1680gagccgctgg tcgaccaggt cctgcaggcc ttcgatcccg aggagcaggc ggccgccctg 1740cggcagctgg cacgaccctg a 176110586PRTC. necator 10Met Pro Phe Ala Leu His Gly Ile Pro Val Ser Arg Gly Val Ala Ile1 5 10 15Gly Arg Ala His Leu Leu Ala Pro Ala Ala Leu Asp Val Ser His Tyr 20 25 30Leu Val Asp Glu Asp Gln Leu Asp Ala Glu Val Glu Arg Leu Arg Ala 35 40 45Ala Arg Ala Ala Val Arg Ala Glu Leu Ala Ala Leu Lys Arg Asp Leu 50 55 60Pro Arg Asp Ala Pro Glu Glu Leu Gly Ala Phe Leu Asp Val His Ala65 70 75 80Met Ile Leu Asp Asp Glu Ala Leu Ala Arg Glu Pro Glu Ala Leu Ile 85 90 95Arg Gly Arg Arg Tyr Asn Ala Glu Trp Ala Leu Thr Thr Arg Leu Glu 100 105 110Glu Leu Met Arg Gln Phe Asp Glu Ile Glu Asp Glu Tyr Leu Arg Glu 115 120 125Arg Lys Thr Asp Ile Arg Gln Val Val Glu Arg Ile Leu Lys Ala Leu 130 135 140Ala Gly Ala Pro Val Leu Val Pro Ala Pro Val Pro Ala Leu Ala Ala145 150 155 160Asp Gly Glu Ala Ala Thr Gly Val Ile Val Val Ala His Asp Ile Ala 165 170 175Pro Ala Asp Met Leu Gln Phe Arg His Thr Val Phe His Gly Phe Val 180 185 190Thr Asp Met Gly Gly Arg Thr Ser His Thr Ala Ile Val Ala Arg Ser 195 200 205Leu Asp Ile Pro Ala Ala Val Gly Val Gln Ser Ala Ser Glu Leu Ile 210 215 220Arg Gln Asp Asp Trp Ile Ile Ile Asp Gly Asp Ala Gly Leu Val Ile225 230 235 240Val Asp Pro Thr Ala Ile Ile Leu Glu Glu Tyr Arg His Arg Gln Ser 245 250 255Glu Arg Ala Leu Glu Lys Lys Arg Leu Gln Arg Leu Arg His Thr Pro 260 265 270Ala Val Thr Leu Asp Gly Leu Glu Ile Asp Leu Leu Ala Asn Ile Glu 275 280 285Met Ala Glu Asp Ala Gly Ala Ala Leu Ala Ala Gly Ala Val Gly Val 290 295 300Gly Leu Phe Arg Ser Glu Phe Leu Phe Met Asn Arg Arg Asp Glu Leu305 310 315 320Pro Gly Glu Asp Glu Gln Phe Gln Ala Tyr Arg Gly Ala Val Asp Ala 325 330 335Met His Gly Leu Pro Val Thr Ile Arg Thr Ile Asp Ile Gly Ala Asp 340 345 350Lys Pro Leu Asp Ala Arg Gly Asp Glu Phe Glu Thr Ala Leu Asn Pro 355 360 365Ala Leu Gly Leu Arg Ala Ile Arg Trp Ser Leu Ser Glu Pro Gly Met 370 375 380Phe Leu Thr Gln Leu Arg Ala Leu Leu Arg Ala Ser Ala Phe Gly Pro385 390 395 400Val Arg Leu Leu Val Pro Met Leu Ala His Ala Ser Glu Ile Asp Gln 405 410 415Thr Leu Ala Leu Ile Ala Lys Ala Lys Arg Gln Leu Asp Glu Arg Gly 420 425 430Glu Ala Tyr Asp Pro Gly Met Lys Val Gly Ala Met Ile Glu Ile Pro 435 440 445Ala Ala Val Leu Leu Leu Pro Leu Phe Leu Arg Lys Met Asp Phe Leu 450 455 460Ser Ile Gly Thr Asn Asp Leu Ile Gln Tyr Thr Leu Ala Ile Asp Arg465 470 475 480Ala Asp Asn Ala Val Ala His Leu Phe Asp Pro Leu His Pro Ala Val 485 490 495Leu Gln Leu Val Ala Arg Thr Ile Arg Glu Ala Asn Arg Ala Gly Val 500 505 510Pro Val Ala Val Cys Gly Glu Met Ala Gly Asp Pro Ser Met Thr Arg 515 520 525Leu Leu Leu Gly Met Gly Leu Arg Glu Phe Ser Met His Pro Ala Gln 530 535 540Leu Leu Arg Val Lys Gln Glu Ile Leu His Ala His Cys Glu Arg Leu545 550 555 560Glu Pro Leu Val Asp Gln Val Leu Gln Ala Phe Asp Pro Glu Glu Gln 565 570 575Ala Ala Ala Leu Arg Gln Leu Ala Arg Pro 580 58511972DNAC. necator 11atggaactca ccggcgtcac ctcccagtcc atcttcgacg acaacgcagc cgacatcaaa 60ctctcgtggg tggccggcct ggaaggtgcg gatcgcgcct tcgatgtgga gttcgcccgc 120gaagccacct ccgccgccga cctggtgggc cacctgaacc tgatccaccc caaccgcatc 180caggtgctcg gcaagcccga gatcacttat taccagcgac tggacgacga gacccgcaag 240cgccagatgg gcgagctgat cctgctggag ccgcccttcc tggtgatcgc cgacggcatg 300gagccgccgc ccgacctgga actgcgctgc acgcgctcgt ccacgccgct gttcaccacg 360ccggtgtcgt cggccgcggt gatcgaccac ctgcgcctgt acctgtcgcg tatctccgcg 420ccgcgcgtga ccatgcacgg ggtattcctc gacatcctgg gcatgggcgt gctgatcatg 480ggcgaatcgg gcctgggcaa gagcgaactg ggcctggaac tgatctcgcg cggccacggg 540ctggtggccg atgatgccgt ggacttcgtg cgcctggggc cggatttcat tgaaggccgc 600tgcccgccgc tgctgcagaa cctgcttgaa gtacgcggtc tgggcctgct cgacatcaag 660accatcttcg gtgagaccgc ggtgcgccgg aagatgaaga tcaagctggt ggtgcagttg 720gtgcgtcgca atgacggcga gttcgagcgg ctgccgctcg attcgcaata cctcgacgtg 780ctgggcctgc cgatccacat ggtcaagatc caggtggcgg ccgggcgcaa cctggccgtg 840ctggtcgagg ccgcggtgcg caacaccatc ctgcgcctgc gcggcatcga tacgctgcgc 900gacttcatgg accggcagcg cgccgcgatg caggccgatg cagtatcgcg cggccagggc 960cgcttgctct ga 97212323PRTC. necator 12Met Glu Leu Thr Gly Val Thr Ser Gln Ser Ile Phe Asp Asp Asn Ala1 5 10 15Ala Asp Ile Lys Leu Ser Trp Val Ala Gly Leu Glu Gly Ala Asp Arg 20 25 30Ala Phe Asp Val Glu Phe Ala Arg Glu Ala Thr Ser Ala Ala Asp Leu 35 40 45Val Gly His Leu Asn Leu Ile His Pro Asn Arg Ile Gln Val Leu Gly 50 55 60Lys Pro Glu Ile Thr Tyr Tyr Gln Arg Leu Asp Asp Glu Thr Arg Lys65 70 75 80Arg Gln Met Gly Glu Leu Ile Leu Leu Glu Pro Pro Phe Leu Val Ile 85 90 95Ala Asp Gly Met Glu Pro Pro Pro Asp Leu Glu Leu Arg Cys Thr Arg 100 105 110Ser Ser Thr Pro Leu Phe Thr Thr Pro Val Ser Ser Ala Ala Val Ile 115 120 125Asp His Leu Arg Leu Tyr Leu Ser Arg Ile Ser Ala Pro Arg Val Thr 130 135 140Met His Gly Val Phe Leu Asp Ile Leu Gly Met Gly Val Leu Ile Met145 150 155 160Gly Glu Ser Gly Leu Gly Lys Ser Glu Leu Gly Leu Glu Leu Ile Ser 165 170 175Arg Gly His Gly Leu Val Ala Asp Asp Ala Val Asp Phe Val Arg Leu 180 185 190Gly Pro Asp Phe Ile Glu Gly Arg Cys Pro Pro Leu Leu Gln Asn Leu 195 200 205Leu Glu Val Arg Gly Leu Gly Leu Leu Asp Ile Lys Thr Ile Phe Gly 210 215 220Glu Thr Ala Val Arg Arg Lys Met Lys Ile Lys Leu Val Val Gln Leu225 230 235 240Val Arg Arg Asn Asp Gly Glu Phe Glu Arg Leu Pro Leu Asp Ser Gln 245 250 255Tyr Leu Asp Val Leu Gly Leu Pro Ile His Met Val Lys Ile Gln Val 260 265 270Ala Ala Gly Arg Asn Leu Ala Val Leu Val Glu Ala Ala Val Arg Asn 275 280 285Thr Ile Leu Arg Leu Arg Gly Ile Asp Thr Leu Arg Asp Phe Met Asp 290 295 300Arg Gln Arg Ala Ala Met Gln Ala Asp Ala Val Ser Arg Gly Gln Gly305 310 315 320Arg Leu Leu13456DNAC. necator 13atgaatcgtt tggccaaatt gctgccaccc ggcaacatca ccctcgacgt cagcgtcacc 60agcaagaagc gtgtgttcga gcaggccggg ctcctcttcg agaacaacca tggtgtggcg 120cgcgccatcg tgacggacaa cctgttcgcg cgcgagtcgc ttggatccac cggcctgggc 180gccggcgtgg caatcccgca cggccgcatc aagggcctga agcagccgct ggccgcgttc 240atgcgcctgg ccgaaccaat tccgttcgaa tcgcccgatg gcaagccggt atcgctgctg 300atcttcctgc tggtgcccga acaggctacg cagcagcacc tggaaatcct gtccgaaatc 360gcgcaactgc tttccgaccg cgacatgcgt gaagggctgg ccacgctgcc cacgcccgat 420gccgtccatc agttgctgat cgcatggcat ccctga 45614151PRTC. necator 14Met Asn Arg Leu Ala Lys Leu Leu Pro Pro Gly Asn Ile Thr Leu Asp1 5 10 15Val Ser Val Thr Ser Lys Lys Arg Val Phe Glu Gln Ala Gly Leu Leu 20 25 30Phe Glu Asn Asn His Gly Val Ala Arg Ala Ile Val Thr Asp Asn Leu 35 40 45Phe Ala Arg Glu Ser Leu Gly Ser Thr Gly Leu Gly Ala Gly Val Ala 50 55 60Ile Pro His Gly Arg Ile Lys Gly Leu Lys Gln Pro Leu Ala Ala Phe65 70 75 80Met Arg Leu Ala Glu Pro Ile Pro Phe Glu Ser Pro Asp Gly Lys Pro 85 90 95Val Ser Leu Leu Ile Phe Leu Leu Val Pro Glu Gln Ala Thr Gln Gln 100 105 110His Leu Glu Ile Leu Ser Glu Ile Ala Gln Leu Leu Ser Asp Arg Asp 115 120 125Met Arg Glu Gly Leu Ala Thr Leu Pro Thr Pro Asp Ala Val His Gln 130 135 140Leu Leu Ile Ala Trp His Pro145 15015351DNAC. necator 15atgaacttca agatcagtgg acaccacctg gacatcacgc cccctctgcg tgagtacgtg 60gaaacgaagc tggagcgaat cgtcaggcat ttcgatcaag tcattggcgt tagtgtgctg 120ctctctgtcg acaaccacaa ggaaaaggac cggcgtcagt acgcggaaat caatctacat 180ctcaagggca aggacatctt tgtcgaagcg catcacgaag acctgtatgc agcgatcgac 240gcactcgtcg acaagctgga ccgtcaggtg atccgctaca aggatcgcgt gcaaggccac 300gaccgcgaag cggtcaagta ccagatggcc gcagcgcaaa tgcagcaatg a 35116116PRTC. necator 16Met Asn Phe Lys Ile Ser Gly His His Leu Asp Ile Thr Pro Pro Leu1 5 10 15Arg Glu Tyr Val Glu Thr Lys Leu Glu Arg Ile Val Arg His Phe Asp 20 25 30Gln Val Ile Gly Val Ser Val Leu Leu Ser Val Asp Asn His Lys Glu 35 40 45Lys Asp Arg Arg Gln Tyr Ala Glu Ile Asn Leu His Leu Lys Gly Lys 50 55 60Asp Ile Phe Val Glu Ala His His Glu Asp Leu Tyr Ala Ala Ile Asp65 70 75 80Ala Leu Val Asp Lys Leu Asp Arg Gln Val Ile Arg Tyr Lys Asp Arg 85 90 95Val Gln Gly His Asp Arg Glu Ala Val Lys Tyr Gln Met Ala Ala Ala 100 105 110Gln Met Gln Gln 115171482DNAC. necator 17atgaaaccgt cgctacagct ccgcctttcc cagcatctgg ccctgacccc gcaactgcag 60cagtcgatcc ggctgctgca gctttccacg ctggaactgc agcaagaggt cgaacaggca 120ctgacggaaa accctctgct cgaacgcgag aacgactgga tcgaaagccc gctgcgcgtg 180gcggccgacg gctcggtcaa cctgcagagc gcaccggcgc ccgcgccggc agagccgcag 240ggcaatggcg aggcccgcgc cgacggcgcg gctgacgacg acagctatgg cgacagcggc 300aacggcgacg actatggcag cagcgactgg agcctggacg actttgcccg ccgcccccag 360ggcgacgagg acgaaaaaac gccgatgcag ctgcgcgaag ccgagcccac gctgcgcgag 420tacctgatgg aacagctcac gccgctgaag

atctcggcgc gcgacaaggg cctggccatc 480ttcctgatcg aatcgctcga cgacgatggc tacctgagcg catcgcttga ggagatctgc 540acggagttgc cggaagaact cgagttcgag atcgaggagg tccacgccat cctcacgctg 600ctgcagagct tcgacccgcc cggcgtgggc gcgcgcaacg ccgccgagtg cctggccctg 660cagttgcggc gcctgacgca cccgcagcgc gaactggccc tgaacatcgt gaccaaccac 720ctggagttgc tcgcagtacg cgactacacg cggctgaaga aggcgctgca ggtggatgaa 780gcggcgctga agtccgcaca cgaactgatc cgctcgctgg cgccctaccc cggccatgca 840tacagccgcc cggaggcgga cttcgtggtg ccggacgtgt tcgtgcgcaa gggtggcggc 900ggctggatcg cgcagctcaa tccggatgtg atgccgaggc tgcgcatcaa tgacatgtat 960gcgcaaatcc tacgtggcgc aaagggtgag tccggcaccg ccgggctgca gcagaagctg 1020caagaggcgc gctggctgat caagaacatc cagcagaggt tcgacaaaat cctgcgtgtc 1080tcgcaggcca ttgtcgagcg tcaaaagaac tttttcagcc acggcgaaat cgccatgcgc 1140cccttggttt tgcgggagat tgccgataca ctgggtttac acgagtcaac catctcccgg 1200gtgacgacca ataaatatat ggcaacgccg atgggtactt tcgaactgaa gtacttcttc 1260ggcagccacg tgtccaccga aaccggtggc gcggcttcgt caacggccat ccgcgccttg 1320atcaagcaac tgataggagc cgaagacccg aggaatcccc tttccgacag tcgcattgcc 1380gaactgctgg gcgaacaagg cttcgttgtc gcacgccgca ccgttgccaa gtatcgcgaa 1440gccctgaaga tccccgcagt caatctccgc aagtctttgt ag 148218493PRTC. necator 18Met Lys Pro Ser Leu Gln Leu Arg Leu Ser Gln His Leu Ala Leu Thr1 5 10 15Pro Gln Leu Gln Gln Ser Ile Arg Leu Leu Gln Leu Ser Thr Leu Glu 20 25 30Leu Gln Gln Glu Val Glu Gln Ala Leu Thr Glu Asn Pro Leu Leu Glu 35 40 45Arg Glu Asn Asp Trp Ile Glu Ser Pro Leu Arg Val Ala Ala Asp Gly 50 55 60Ser Val Asn Leu Gln Ser Ala Pro Ala Pro Ala Pro Ala Glu Pro Gln65 70 75 80Gly Asn Gly Glu Ala Arg Ala Asp Gly Ala Ala Asp Asp Asp Ser Tyr 85 90 95Gly Asp Ser Gly Asn Gly Asp Asp Tyr Gly Ser Ser Asp Trp Ser Leu 100 105 110Asp Asp Phe Ala Arg Arg Pro Gln Gly Asp Glu Asp Glu Lys Thr Pro 115 120 125Met Gln Leu Arg Glu Ala Glu Pro Thr Leu Arg Glu Tyr Leu Met Glu 130 135 140Gln Leu Thr Pro Leu Lys Ile Ser Ala Arg Asp Lys Gly Leu Ala Ile145 150 155 160Phe Leu Ile Glu Ser Leu Asp Asp Asp Gly Tyr Leu Ser Ala Ser Leu 165 170 175Glu Glu Ile Cys Thr Glu Leu Pro Glu Glu Leu Glu Phe Glu Ile Glu 180 185 190Glu Val His Ala Ile Leu Thr Leu Leu Gln Ser Phe Asp Pro Pro Gly 195 200 205Val Gly Ala Arg Asn Ala Ala Glu Cys Leu Ala Leu Gln Leu Arg Arg 210 215 220Leu Thr His Pro Gln Arg Glu Leu Ala Leu Asn Ile Val Thr Asn His225 230 235 240Leu Glu Leu Leu Ala Val Arg Asp Tyr Thr Arg Leu Lys Lys Ala Leu 245 250 255Gln Val Asp Glu Ala Ala Leu Lys Ser Ala His Glu Leu Ile Arg Ser 260 265 270Leu Ala Pro Tyr Pro Gly His Ala Tyr Ser Arg Pro Glu Ala Asp Phe 275 280 285Val Val Pro Asp Val Phe Val Arg Lys Gly Gly Gly Gly Trp Ile Ala 290 295 300Gln Leu Asn Pro Asp Val Met Pro Arg Leu Arg Ile Asn Asp Met Tyr305 310 315 320Ala Gln Ile Leu Arg Gly Ala Lys Gly Glu Ser Gly Thr Ala Gly Leu 325 330 335Gln Gln Lys Leu Gln Glu Ala Arg Trp Leu Ile Lys Asn Ile Gln Gln 340 345 350Arg Phe Asp Lys Ile Leu Arg Val Ser Gln Ala Ile Val Glu Arg Gln 355 360 365Lys Asn Phe Phe Ser His Gly Glu Ile Ala Met Arg Pro Leu Val Leu 370 375 380Arg Glu Ile Ala Asp Thr Leu Gly Leu His Glu Ser Thr Ile Ser Arg385 390 395 400Val Thr Thr Asn Lys Tyr Met Ala Thr Pro Met Gly Thr Phe Glu Leu 405 410 415Lys Tyr Phe Phe Gly Ser His Val Ser Thr Glu Thr Gly Gly Ala Ala 420 425 430Ser Ser Thr Ala Ile Arg Ala Leu Ile Lys Gln Leu Ile Gly Ala Glu 435 440 445Asp Pro Arg Asn Pro Leu Ser Asp Ser Arg Ile Ala Glu Leu Leu Gly 450 455 460Glu Gln Gly Phe Val Val Ala Arg Arg Thr Val Ala Lys Tyr Arg Glu465 470 475 480Ala Leu Lys Ile Pro Ala Val Asn Leu Arg Lys Ser Leu 485 49019591DNAC. necator 19atgcgcaagg gtgaaatgac gcgcgtggcg attctggatg ccgcactgga attgtcgtcc 60cgcgacgggc tcgaaggtct gaccatcggg ctgctcgcgg aacgcatgca gatgagcaag 120agcggtgtct tcgcgcattt cggttcgcgc gaagacctgc aggtggaggt ggtgcgggag 180tatcaccgcc ggttcgagca ggaggtgttc tatccctcgc tgcaggagcc gcgcggcctg 240ccccggctat ggtcgatggt gcggcgctgg atggagaagc gcatccagga agtgacgact 300ggatgcatct acatcagcgg cgccgtggag tacgacgacc gtgccggcag cctggtgcgt 360gacgagctgg tcaagagcgt caccatctgg cgggcagcgc tcacgcgcgc catcaaccag 420gcgcgggagg aagggcacct gcgcgcggac tgcgatccgc gcctgatgct gttcgagatg 480tacagccttg aactaggctt gcatcatgac gcccgtttcc tgcgcctgcc tgacagtgcc 540gagcttgcca tggtcgcgct caacaagctg attcagtctt accgtacctg a 59120196PRTC. necator 20Met Arg Lys Gly Glu Met Thr Arg Val Ala Ile Leu Asp Ala Ala Leu1 5 10 15Glu Leu Ser Ser Arg Asp Gly Leu Glu Gly Leu Thr Ile Gly Leu Leu 20 25 30Ala Glu Arg Met Gln Met Ser Lys Ser Gly Val Phe Ala His Phe Gly 35 40 45Ser Arg Glu Asp Leu Gln Val Glu Val Val Arg Glu Tyr His Arg Arg 50 55 60Phe Glu Gln Glu Val Phe Tyr Pro Ser Leu Gln Glu Pro Arg Gly Leu65 70 75 80Pro Arg Leu Trp Ser Met Val Arg Arg Trp Met Glu Lys Arg Ile Gln 85 90 95Glu Val Thr Thr Gly Cys Ile Tyr Ile Ser Gly Ala Val Glu Tyr Asp 100 105 110Asp Arg Ala Gly Ser Leu Val Arg Asp Glu Leu Val Lys Ser Val Thr 115 120 125Ile Trp Arg Ala Ala Leu Thr Arg Ala Ile Asn Gln Ala Arg Glu Glu 130 135 140Gly His Leu Arg Ala Asp Cys Asp Pro Arg Leu Met Leu Phe Glu Met145 150 155 160Tyr Ser Leu Glu Leu Gly Leu His His Asp Ala Arg Phe Leu Arg Leu 165 170 175Pro Asp Ser Ala Glu Leu Ala Met Val Ala Leu Asn Lys Leu Ile Gln 180 185 190Ser Tyr Arg Thr 195211308DNAC. necator 21atgtcttccg cagcaccgac caatattgct ggccaaaagc acgcactccc gtcttacctc 60aatgccgacc atctcggccc ctggggcatc tacctgcagc aggtcgaccg tgtcacgcct 120tacctgggct cgctggcacg ctgggtcgaa accctcaagc gccccaagcg cgcgatgatc 180gtcgacgttc ccatcgaact ggataacggc accattgccc atttcgaggg ctatcgggtg 240cagcacaacc tgtcgcgcgg cccgggcaag ggcggcgtgc gcttccacca ggacgtgacc 300ctgtccgagg tgatggcgct gtcggcctgg atgtcggtga agaatgccgc ggtcaacgtg 360ccctacggcg gtgccaaggg cggcatccgc gtcgatccgc gcacgctctc gcacgccgag 420ctggaacgcc tgacgcgccg ctacaccagc gaaatcaaca tcatcatcgg gccgagcaag 480gatattccgg cgccggacgt gaacaccaac gcccaggtca tggcctggat gatggacacg 540tactcgatga actccggcag cacggccacc ggcgtggtga ccggcaagcc gatctcgctg 600ggcggctcgc tcggccgcca cgaagccacc ggccgcggcg tgttcgtggt cggctccgag 660gccgcgcgca atatcggcct ggagatcaag ggcgcgcgcg tggcggtgca gggcttcggc 720aacgtgggcg cggtggcggc caagctgttc catgaggccg gcgccaaggt ggtggcggtg 780caggaccacc gcaccacgct gttcgacccg gccggcctgg acgtgccggc gatgatggaa 840tacgcctcgc acagcggcac catcgaaggc ttccgcggcg aagtcctgcg caccgagcag 900ttctgggaag tcgactgcga catcctgatc ccggccgcgc tggaaggcca gatcacggtg 960cagaacgcgc ccaagatcac ggcaaagctg gtgattgaag gcgccaacgg cccgaccacg 1020ccgcaagccg acgatatcct gcgcgagcgc aatatcctgg tctgtcccga cgtgatcgcc 1080aacgccggcg gcgtgaccgt gtcctacttc gaatgggtgc aggatttctc cagctttttc 1140tggaccgagg aagaaatcaa ccagcgcctg gtacggatca tgcaagaagc cttccgggca 1200atctggcaag tggcacagga caacaaggtg acgctgcgca cggcggcgtt tatcgtggcc 1260tgtacgcgga tcctgcaggc gcgcgagatg cgcggcctgt atccctga 130822435PRTC. necator 22Met Ser Ser Ala Ala Pro Thr Asn Ile Ala Gly Gln Lys His Ala Leu1 5 10 15Pro Ser Tyr Leu Asn Ala Asp His Leu Gly Pro Trp Gly Ile Tyr Leu 20 25 30Gln Gln Val Asp Arg Val Thr Pro Tyr Leu Gly Ser Leu Ala Arg Trp 35 40 45Val Glu Thr Leu Lys Arg Pro Lys Arg Ala Met Ile Val Asp Val Pro 50 55 60Ile Glu Leu Asp Asn Gly Thr Ile Ala His Phe Glu Gly Tyr Arg Val65 70 75 80Gln His Asn Leu Ser Arg Gly Pro Gly Lys Gly Gly Val Arg Phe His 85 90 95Gln Asp Val Thr Leu Ser Glu Val Met Ala Leu Ser Ala Trp Met Ser 100 105 110Val Lys Asn Ala Ala Val Asn Val Pro Tyr Gly Gly Ala Lys Gly Gly 115 120 125Ile Arg Val Asp Pro Arg Thr Leu Ser His Ala Glu Leu Glu Arg Leu 130 135 140Thr Arg Arg Tyr Thr Ser Glu Ile Asn Ile Ile Ile Gly Pro Ser Lys145 150 155 160Asp Ile Pro Ala Pro Asp Val Asn Thr Asn Ala Gln Val Met Ala Trp 165 170 175Met Met Asp Thr Tyr Ser Met Asn Ser Gly Ser Thr Ala Thr Gly Val 180 185 190Val Thr Gly Lys Pro Ile Ser Leu Gly Gly Ser Leu Gly Arg His Glu 195 200 205Ala Thr Gly Arg Gly Val Phe Val Val Gly Ser Glu Ala Ala Arg Asn 210 215 220Ile Gly Leu Glu Ile Lys Gly Ala Arg Val Ala Val Gln Gly Phe Gly225 230 235 240Asn Val Gly Ala Val Ala Ala Lys Leu Phe His Glu Ala Gly Ala Lys 245 250 255Val Val Ala Val Gln Asp His Arg Thr Thr Leu Phe Asp Pro Ala Gly 260 265 270Leu Asp Val Pro Ala Met Met Glu Tyr Ala Ser His Ser Gly Thr Ile 275 280 285Glu Gly Phe Arg Gly Glu Val Leu Arg Thr Glu Gln Phe Trp Glu Val 290 295 300Asp Cys Asp Ile Leu Ile Pro Ala Ala Leu Glu Gly Gln Ile Thr Val305 310 315 320Gln Asn Ala Pro Lys Ile Thr Ala Lys Leu Val Ile Glu Gly Ala Asn 325 330 335Gly Pro Thr Thr Pro Gln Ala Asp Asp Ile Leu Arg Glu Arg Asn Ile 340 345 350Leu Val Cys Pro Asp Val Ile Ala Asn Ala Gly Gly Val Thr Val Ser 355 360 365Tyr Phe Glu Trp Val Gln Asp Phe Ser Ser Phe Phe Trp Thr Glu Glu 370 375 380Glu Ile Asn Gln Arg Leu Val Arg Ile Met Gln Glu Ala Phe Arg Ala385 390 395 400Ile Trp Gln Val Ala Gln Asp Asn Lys Val Thr Leu Arg Thr Ala Ala 405 410 415Phe Ile Val Ala Cys Thr Arg Ile Leu Gln Ala Arg Glu Met Arg Gly 420 425 430Leu Tyr Pro 43523339DNAC. necator 23atgaagcaga ttaccgccat catcaaaccg ttcaagctcg acgaggtgcg tgaagccctg 60gccgacgtcg gcgtgaccgg tctgacggtg accgaagtga agggatttgg ccgccagaaa 120gggcataccg agctctaccg tggcgccgag tacgtggtcg acttcctgcc caagatcaag 180atcgaagtgg tggtggccga gaaccagctg gacaccgtgc tggacgccat cgtcaaggcc 240gcccacaccg gcaagatcgg cgacggcaag atcttcgtca ccgagatcga gcgtgtgatc 300cgcatccgca ccggcgagca ggacgaagcc gcggtctga 33924112PRTC. necator 24Met Lys Gln Ile Thr Ala Ile Ile Lys Pro Phe Lys Leu Asp Glu Val1 5 10 15Arg Glu Ala Leu Ala Asp Val Gly Val Thr Gly Leu Thr Val Thr Glu 20 25 30Val Lys Gly Phe Gly Arg Gln Lys Gly His Thr Glu Leu Tyr Arg Gly 35 40 45Ala Glu Tyr Val Val Asp Phe Leu Pro Lys Ile Lys Ile Glu Val Val 50 55 60Val Ala Glu Asn Gln Leu Asp Thr Val Leu Asp Ala Ile Val Lys Ala65 70 75 80Ala His Thr Gly Lys Ile Gly Asp Gly Lys Ile Phe Val Thr Glu Ile 85 90 95Glu Arg Val Ile Arg Ile Arg Thr Gly Glu Gln Asp Glu Ala Ala Val 100 105 110252412DNAC. necator 25gtgaacgcgc gcaccggcgc tcccaatctt cccgatccgc tcggcgacga tgtcgtcggc 60gagcgtgccg tcgcgcccgc gctggcgccg ggcaagtcca ggggcgcggc gctgcaggac 120gagctggccg cgatgcagga tgtggcgccg gcggctgaca gcctgtttat cgacgcagtg 180ctggcgcagt cataccggca cttcttcggc ccgacctcgc agccggcggt gccgccgcgc 240cagcaggtca tctccatcac ccggctgatg gagaaactcg cgtacctgaa ggcgcccgac 300ctggcgcgcg tgcgcgaggc cttccagttt tccgacgaag cccacctggg ccagtaccgc 360cagagcggtg agccctacat cacccatccg gtggcggtgg ccgagctgtg cgcggactgg 420aagctggatg tgcagtccat catggcggcg ctgctgcacg acgtgatgga agaccagggc 480atcaccaaga gcgagctggt cgagaaattc ggccccaagg tcgccgaact ggtcgacggc 540ctgaccaaat tggacaagct cgaattccag agccgcgagc aggcgcaggc ggagagcttc 600cgcaagatgc tgctggcgat ggcgcgcgac gtgcgcgtga tcctggtgaa gctggccgac 660cgtacgcaca acatgcgcac gctcgacttc gtcccgccgg agaagcgccg ccgcatcgcg 720ctggagacca tggagatcta tgcgccgatc gcgcaccgtc tcggtctcaa cacgatctac 780cgcgagctgc aggagctgtc cttcaaggtc ggctcgccgt tccgctacgc cacgctggaa 840aaagccgtca aggccgcgcg cggcaaccgc cgcgaggtgg tcaagcgtat cctggaagcc 900gcgcagaagg ggctggccga cgccggcatc gtggccgaac tgtccgggcg cgagaaaacg 960ctctacagca tctaccgcaa gatgcacgac aagcagctgt cgttctcgca ggtgctggac 1020gtatatggtt tccgcgtggt ggtggaaacg cagatgcact gctacatggc gatgggcgcg 1080ctgcatgggc tgtacaagcc catgcccggc aagttcaagg actacatcgc catccccaag 1140atcaacggct accagtcgct gcacaccacg ctggtgggtc cgttcggcac gccggtggag 1200ttccagatcc gcacgcgcga catgcaccag atcgccgagg ccggcgtggc cgcgcactgg 1260atgtacaagc accaggccga tcacgccaac gatatccagc agcaggcgca ccagtggctg 1320cagtcgctgc tcgatatcca gagccagacc ggcgattcgc aggaattcct cgagcacgtc 1380aagatcgacc tgttcccgga tgcggtctac gtgttcacgc ccaagggcca tatccgcgcg 1440ctgccgcgcg gcgccaccgc gctggacttt gcctacgcgg tgcacagcga cctgggcaac 1500cagtgcgtcg cggtcaagat caacaacgag atgttgccgc tgcgcaccga gctcaagagc 1560ggcgatatcg tcgaggtggt gacggcgccg tactccaagc ccaatccggc gtggctgtcg 1620ttcgtgcgca ccggcaaggc gcgcgcggcg atccgccact acctgaagac caccaagctc 1680gacgaagcca tccagcttgg cgagcgcctg ctggaacagt cggcgcgcca gctcggcttc 1740gagctcaagg cggtgccgca gtcggtgtgg gaccgcatgg tgcagtggac cggcaacaag 1800cagcgcgaag acatctttgc cgacctggca ctgggccggc gcgtgccggc ggtggtggcc 1860aagcgcatgg agatcctgct ccaggagctg tccggcgatg tcgacagcgc gctgctggcg 1920gcggtgcaga ccttcgccgg cgaagaagcg cccgcggtgc cgatcaccgg cgacgaaggc 1980atgtcgatga tcttctcggc gtgctgccgc ccgatcccgg gcgactccat cgttggctac 2040ctgggcaagg gcgaagggct gcagatccac gtgcaggact gcaagatcgc caagcgcctg 2100cacagcaagg atccggagca ctggatcgac gtgatgtggg ccaagaagac cacgcgcgcc 2160ttcgacgtgt cgatcaaggt gatggtgcgc aacgtcaagg gcatcgttgc gcgcgtggct 2220gccgacctga ccgccgccga cgccaacgtc gcgcacgtgg ccatggagca gcaggacgcc 2280ggccaccagg aagccaccta tatgcagttt atcatccagg tgcagaaccg cctgcacctg 2340gccaacgtga tgcgcgggct gcgccgcaac ccggacgtca tccggatatt ccgcgaccgc 2400aacgacggct ag 241226803PRTC. necator 26Met Asn Ala Arg Thr Gly Ala Pro Asn Leu Pro Asp Pro Leu Gly Asp1 5 10 15Asp Val Val Gly Glu Arg Ala Val Ala Pro Ala Leu Ala Pro Gly Lys 20 25 30Ser Arg Gly Ala Ala Leu Gln Asp Glu Leu Ala Ala Met Gln Asp Val 35 40 45Ala Pro Ala Ala Asp Ser Leu Phe Ile Asp Ala Val Leu Ala Gln Ser 50 55 60Tyr Arg His Phe Phe Gly Pro Thr Ser Gln Pro Ala Val Pro Pro Arg65 70 75 80Gln Gln Val Ile Ser Ile Thr Arg Leu Met Glu Lys Leu Ala Tyr Leu 85 90 95Lys Ala Pro Asp Leu Ala Arg Val Arg Glu Ala Phe Gln Phe Ser Asp 100 105 110Glu Ala His Leu Gly Gln Tyr Arg Gln Ser Gly Glu Pro Tyr Ile Thr 115 120 125His Pro Val Ala Val Ala Glu Leu Cys Ala Asp Trp Lys Leu Asp Val 130 135 140Gln Ser Ile Met Ala Ala Leu Leu His Asp Val Met Glu Asp Gln Gly145 150 155 160Ile Thr Lys Ser Glu Leu Val Glu Lys Phe Gly Pro Lys Val Ala Glu 165 170 175Leu Val Asp Gly Leu Thr Lys Leu Asp Lys Leu Glu Phe Gln Ser Arg 180 185 190Glu Gln Ala Gln Ala Glu Ser Phe Arg Lys Met Leu Leu Ala Met Ala 195 200 205Arg Asp Val Arg Val Ile Leu Val Lys Leu Ala Asp Arg Thr His Asn 210 215 220Met Arg Thr Leu Asp Phe Val Pro Pro Glu Lys Arg Arg Arg Ile Ala225 230 235 240Leu Glu Thr Met Glu Ile Tyr Ala Pro Ile Ala His Arg Leu Gly Leu 245 250 255Asn Thr Ile Tyr Arg Glu Leu Gln Glu Leu Ser Phe

Lys Val Gly Ser 260 265 270Pro Phe Arg Tyr Ala Thr Leu Glu Lys Ala Val Lys Ala Ala Arg Gly 275 280 285Asn Arg Arg Glu Val Val Lys Arg Ile Leu Glu Ala Ala Gln Lys Gly 290 295 300Leu Ala Asp Ala Gly Ile Val Ala Glu Leu Ser Gly Arg Glu Lys Thr305 310 315 320Leu Tyr Ser Ile Tyr Arg Lys Met His Asp Lys Gln Leu Ser Phe Ser 325 330 335Gln Val Leu Asp Val Tyr Gly Phe Arg Val Val Val Glu Thr Gln Met 340 345 350His Cys Tyr Met Ala Met Gly Ala Leu His Gly Leu Tyr Lys Pro Met 355 360 365Pro Gly Lys Phe Lys Asp Tyr Ile Ala Ile Pro Lys Ile Asn Gly Tyr 370 375 380Gln Ser Leu His Thr Thr Leu Val Gly Pro Phe Gly Thr Pro Val Glu385 390 395 400Phe Gln Ile Arg Thr Arg Asp Met His Gln Ile Ala Glu Ala Gly Val 405 410 415Ala Ala His Trp Met Tyr Lys His Gln Ala Asp His Ala Asn Asp Ile 420 425 430Gln Gln Gln Ala His Gln Trp Leu Gln Ser Leu Leu Asp Ile Gln Ser 435 440 445Gln Thr Gly Asp Ser Gln Glu Phe Leu Glu His Val Lys Ile Asp Leu 450 455 460Phe Pro Asp Ala Val Tyr Val Phe Thr Pro Lys Gly His Ile Arg Ala465 470 475 480Leu Pro Arg Gly Ala Thr Ala Leu Asp Phe Ala Tyr Ala Val His Ser 485 490 495Asp Leu Gly Asn Gln Cys Val Ala Val Lys Ile Asn Asn Glu Met Leu 500 505 510Pro Leu Arg Thr Glu Leu Lys Ser Gly Asp Ile Val Glu Val Val Thr 515 520 525Ala Pro Tyr Ser Lys Pro Asn Pro Ala Trp Leu Ser Phe Val Arg Thr 530 535 540Gly Lys Ala Arg Ala Ala Ile Arg His Tyr Leu Lys Thr Thr Lys Leu545 550 555 560Asp Glu Ala Ile Gln Leu Gly Glu Arg Leu Leu Glu Gln Ser Ala Arg 565 570 575Gln Leu Gly Phe Glu Leu Lys Ala Val Pro Gln Ser Val Trp Asp Arg 580 585 590Met Val Gln Trp Thr Gly Asn Lys Gln Arg Glu Asp Ile Phe Ala Asp 595 600 605Leu Ala Leu Gly Arg Arg Val Pro Ala Val Val Ala Lys Arg Met Glu 610 615 620Ile Leu Leu Gln Glu Leu Ser Gly Asp Val Asp Ser Ala Leu Leu Ala625 630 635 640Ala Val Gln Thr Phe Ala Gly Glu Glu Ala Pro Ala Val Pro Ile Thr 645 650 655Gly Asp Glu Gly Met Ser Met Ile Phe Ser Ala Cys Cys Arg Pro Ile 660 665 670Pro Gly Asp Ser Ile Val Gly Tyr Leu Gly Lys Gly Glu Gly Leu Gln 675 680 685Ile His Val Gln Asp Cys Lys Ile Ala Lys Arg Leu His Ser Lys Asp 690 695 700Pro Glu His Trp Ile Asp Val Met Trp Ala Lys Lys Thr Thr Arg Ala705 710 715 720Phe Asp Val Ser Ile Lys Val Met Val Arg Asn Val Lys Gly Ile Val 725 730 735Ala Arg Val Ala Ala Asp Leu Thr Ala Ala Asp Ala Asn Val Ala His 740 745 750Val Ala Met Glu Gln Gln Asp Ala Gly His Gln Glu Ala Thr Tyr Met 755 760 765Gln Phe Ile Ile Gln Val Gln Asn Arg Leu His Leu Ala Asn Val Met 770 775 780Arg Gly Leu Arg Arg Asn Pro Asp Val Ile Arg Ile Phe Arg Asp Arg785 790 795 800Asn Asp Gly27948DNAC. necator 27atgcgcgacc gaatcctagc cgtctacgac acgctccgcc cttctgaacg ccgcctggcc 60gactatgtcg cccggcatgg cgccgccgtg atccggctgt cgatgcccga gctggccgag 120cgcgccggcg tgtcgcagcc caccatcgcg cgcttctgcg cggcgctggg ctacgacggc 180ttccgcgaat tcaagctgca gttcgcgcag aacgttggcg gcggcacgcc cttcgtgcac 240caggacgtcg cggccgacga ccgccccgcg gacatcgccg gcaaggtctt cgaccgcacc 300attgccacgc tgatgagcgt gcgcaatgcg ctgtcggccg accagatcga gcatggcatc 360cagctgctcg ccggcgcgcg ccgcatcgag ttctacggct gcggcaactc cggcatcgtc 420gcgctggata tccagcacaa gttcttccgc ctgggcatgc cgacggttgc gtattccgac 480ccgcacgtgt tcagcatgtc ggccgcgctg ctcgcccgtg gcgacgtggc cgtgctggtc 540tccaacagcg gccgcacctg ggacatgctg accgctgcca cgctggcgcg cagcagcggc 600gccagcgtgc tggcaatcac gcacagcggc tcgccgctgg cgcggctggc tgacgtctgc 660gtgttttccg acgtcgagga agacagcgag gtctacacgc cgatgacctc gcgcatcagc 720cacctggtgc tgggcgacgt cctggccgcc ggcgtggcgc ttgcgcgcgc cgacaccgtc 780gccccccgcc tgcagcgcgc caaggcgcat ctgcgcgaac gacgcattgc cggtgcggag 840ccggcccggc cagtaccgcc ggcacgaaac cgcgccaggc ccgccgcagc cgagccagcc 900acgcccgcgc tgcccgccac ccgcacgcgt cgccgcaagg ccagttga 94828315PRTC. necator 28Met Arg Asp Arg Ile Leu Ala Val Tyr Asp Thr Leu Arg Pro Ser Glu1 5 10 15Arg Arg Leu Ala Asp Tyr Val Ala Arg His Gly Ala Ala Val Ile Arg 20 25 30Leu Ser Met Pro Glu Leu Ala Glu Arg Ala Gly Val Ser Gln Pro Thr 35 40 45Ile Ala Arg Phe Cys Ala Ala Leu Gly Tyr Asp Gly Phe Arg Glu Phe 50 55 60Lys Leu Gln Phe Ala Gln Asn Val Gly Gly Gly Thr Pro Phe Val His65 70 75 80Gln Asp Val Ala Ala Asp Asp Arg Pro Ala Asp Ile Ala Gly Lys Val 85 90 95Phe Asp Arg Thr Ile Ala Thr Leu Met Ser Val Arg Asn Ala Leu Ser 100 105 110Ala Asp Gln Ile Glu His Gly Ile Gln Leu Leu Ala Gly Ala Arg Arg 115 120 125Ile Glu Phe Tyr Gly Cys Gly Asn Ser Gly Ile Val Ala Leu Asp Ile 130 135 140Gln His Lys Phe Phe Arg Leu Gly Met Pro Thr Val Ala Tyr Ser Asp145 150 155 160Pro His Val Phe Ser Met Ser Ala Ala Leu Leu Ala Arg Gly Asp Val 165 170 175Ala Val Leu Val Ser Asn Ser Gly Arg Thr Trp Asp Met Leu Thr Ala 180 185 190Ala Thr Leu Ala Arg Ser Ser Gly Ala Ser Val Leu Ala Ile Thr His 195 200 205Ser Gly Ser Pro Leu Ala Arg Leu Ala Asp Val Cys Val Phe Ser Asp 210 215 220Val Glu Glu Asp Ser Glu Val Tyr Thr Pro Met Thr Ser Arg Ile Ser225 230 235 240His Leu Val Leu Gly Asp Val Leu Ala Ala Gly Val Ala Leu Ala Arg 245 250 255Ala Asp Thr Val Ala Pro Arg Leu Gln Arg Ala Lys Ala His Leu Arg 260 265 270Glu Arg Arg Ile Ala Gly Ala Glu Pro Ala Arg Pro Val Pro Pro Ala 275 280 285Arg Asn Arg Ala Arg Pro Ala Ala Ala Glu Pro Ala Thr Pro Ala Leu 290 295 300Pro Ala Thr Arg Thr Arg Arg Arg Lys Ala Ser305 310 315292229DNAC. necator 29atggtgacgt ccaccgacct gaccggtcgg gtcgcgggca ttccggatgc cgaactggtc 60gaacgcgcac tggcctacgt gcgcgagcat ggtgccgagg tagcgctgcc caccggcgag 120accgtgctgt cgcacgcgca gggcatgctg cgcattctcg acggcctgcg cgtcgacgac 180gccgcgcgtg ccgccgcctg cctgttcggg ctggtggcct tcgtgcccgg taccgaggcc 240gagatcgcac cgcgctttgg agacgaggtg gcacggctgg tcgacggtgt gcggcagctg 300ctgcgcatcg gtgccatcgc cggcagccgc cccgaggccg agccggccgc gccgtccaag 360aacgaagcgc aggcgcgcca cgaacaggtc gaggcgctgc gcaagatgct gctggcgttt 420gcgcaggaca tccgcgtggt gctggtgcgc ctggcctcgc ggctgcagac cctgcgctgg 480ctggccgaga ccaagcaggc gccgcagccg ggcgtggcgc gcgaaacgct ggacatctac 540gcgccgctgg ccaatcgcct tggtatctgg cagatgaagt gggaactgga ggacctggcg 600ttccgcttcg agcagcccga tacctacaag cgcatcgcca aactgctgga cgagaagcgc 660atcgagcgcg aaggctatat cggcggcgcc atcgaacggc tgcagtccga gctggcgact 720gccggcatcc gtgccgaggt cagcgggcgg cccaagcata tctacagcat ctggaagaag 780atgcgcggca aggagctgga ttttgccgac ctgtacgatg tgcgcgcctt ccgcgtgatc 840gtcgacgata tcaaggactg ctacacggtg ctgggtatcg tccaccatat ctggcagccg 900atcccgcgcg agttcgacga ctatatctcg cggcccaagg ccaatggcta caaatcgctg 960catacggtgg tgatcggcga tgacgggcgc gccttcgagg tgcagatccg cacgcatgag 1020atgcaccact ttgccgaata cggcgtggcc gcgcactggc gctacaagga agcgggcagc 1080cgcggctatg ccgggcagtt ctccgccagc gagcgctatg acgagaagat tgcctggctg 1140cgccagctgc tggcgtggaa ggacgatgcc gaccacagcg tggcgcacga tgaatcgtgg 1200gagcagatca agcacgccgc gatcgacgac cacatctacg tgctgacgcc gcaggcgcgc 1260gtggtggcgc tgccgcaggg cgccaccgcg gtggactttg cctactacct gcacagcgat 1320ctcggccacc gctgccgcgg cgcgcgcgtg gacggcacca tggtgccgct gaacacgccg 1380ctgaagaacg gccagaccgt ggagatcatc gcggtcaagc agggcggacc gtcgcgcgac 1440tggctcaacg cggacctggg ctacctggcc agcagccgcg cgcgggccaa ggtgcgggcc 1500tggttcaatg cgctcgattc gcaggagacc atcgcccagg gccgcgtgct gatcgacaag 1560accctgcagc gcgaaggcaa gaccgcggtc aagctggaag acctggccac gcggctgggc 1620ttcaagacgc cggaggacct gtttgcggca gtggccaagg acgagttcag tctgcgccac 1680gtggagcacg cgctgcgaca cccggagggc gaggtccagg cgccgctgag cgaggaagac 1740gctgtcacca agaagagccg cgccaccagc gtggcgcgcg gcgccaagag cggcgtgctg 1800gtggtggggg tggattcgct gatgacgcag atgtcgcgct gctgcaagcc ggcgccgccg 1860gacgacatcg tcggctttgt cacgcgcggg cgcggggtgt cgatccatcg gcgcagctgc 1920cacaccttcc agcaactggc cgggcgcgcg ccggagcggg tgatccagac cgagtggggc 1980cagaagagcc acgccgcggt ctatccggtc gatatccatg tcgaggcgat cgatcgccag 2040gggctgctgc gcgatatctc cgaagtgctg tcgcgcgaga agatcaatgt caccggcgtc 2100aagacgctct ccagcaaggg cgttgcgcgc atgcagttca ctgccgaagt gtccgaggct 2160acgcagctgc agcgcgcgct gcagttgatc gaagacgtcc agggggtgtt gcaggcgaaa 2220agaaagtga 222930742PRTC. necator 30Met Val Thr Ser Thr Asp Leu Thr Gly Arg Val Ala Gly Ile Pro Asp1 5 10 15Ala Glu Leu Val Glu Arg Ala Leu Ala Tyr Val Arg Glu His Gly Ala 20 25 30Glu Val Ala Leu Pro Thr Gly Glu Thr Val Leu Ser His Ala Gln Gly 35 40 45Met Leu Arg Ile Leu Asp Gly Leu Arg Val Asp Asp Ala Ala Arg Ala 50 55 60Ala Ala Cys Leu Phe Gly Leu Val Ala Phe Val Pro Gly Thr Glu Ala65 70 75 80Glu Ile Ala Pro Arg Phe Gly Asp Glu Val Ala Arg Leu Val Asp Gly 85 90 95Val Arg Gln Leu Leu Arg Ile Gly Ala Ile Ala Gly Ser Arg Pro Glu 100 105 110Ala Glu Pro Ala Ala Pro Ser Lys Asn Glu Ala Gln Ala Arg His Glu 115 120 125Gln Val Glu Ala Leu Arg Lys Met Leu Leu Ala Phe Ala Gln Asp Ile 130 135 140Arg Val Val Leu Val Arg Leu Ala Ser Arg Leu Gln Thr Leu Arg Trp145 150 155 160Leu Ala Glu Thr Lys Gln Ala Pro Gln Pro Gly Val Ala Arg Glu Thr 165 170 175Leu Asp Ile Tyr Ala Pro Leu Ala Asn Arg Leu Gly Ile Trp Gln Met 180 185 190Lys Trp Glu Leu Glu Asp Leu Ala Phe Arg Phe Glu Gln Pro Asp Thr 195 200 205Tyr Lys Arg Ile Ala Lys Leu Leu Asp Glu Lys Arg Ile Glu Arg Glu 210 215 220Gly Tyr Ile Gly Gly Ala Ile Glu Arg Leu Gln Ser Glu Leu Ala Thr225 230 235 240Ala Gly Ile Arg Ala Glu Val Ser Gly Arg Pro Lys His Ile Tyr Ser 245 250 255Ile Trp Lys Lys Met Arg Gly Lys Glu Leu Asp Phe Ala Asp Leu Tyr 260 265 270Asp Val Arg Ala Phe Arg Val Ile Val Asp Asp Ile Lys Asp Cys Tyr 275 280 285Thr Val Leu Gly Ile Val His His Ile Trp Gln Pro Ile Pro Arg Glu 290 295 300Phe Asp Asp Tyr Ile Ser Arg Pro Lys Ala Asn Gly Tyr Lys Ser Leu305 310 315 320His Thr Val Val Ile Gly Asp Asp Gly Arg Ala Phe Glu Val Gln Ile 325 330 335Arg Thr His Glu Met His His Phe Ala Glu Tyr Gly Val Ala Ala His 340 345 350Trp Arg Tyr Lys Glu Ala Gly Ser Arg Gly Tyr Ala Gly Gln Phe Ser 355 360 365Ala Ser Glu Arg Tyr Asp Glu Lys Ile Ala Trp Leu Arg Gln Leu Leu 370 375 380Ala Trp Lys Asp Asp Ala Asp His Ser Val Ala His Asp Glu Ser Trp385 390 395 400Glu Gln Ile Lys His Ala Ala Ile Asp Asp His Ile Tyr Val Leu Thr 405 410 415Pro Gln Ala Arg Val Val Ala Leu Pro Gln Gly Ala Thr Ala Val Asp 420 425 430Phe Ala Tyr Tyr Leu His Ser Asp Leu Gly His Arg Cys Arg Gly Ala 435 440 445Arg Val Asp Gly Thr Met Val Pro Leu Asn Thr Pro Leu Lys Asn Gly 450 455 460Gln Thr Val Glu Ile Ile Ala Val Lys Gln Gly Gly Pro Ser Arg Asp465 470 475 480Trp Leu Asn Ala Asp Leu Gly Tyr Leu Ala Ser Ser Arg Ala Arg Ala 485 490 495Lys Val Arg Ala Trp Phe Asn Ala Leu Asp Ser Gln Glu Thr Ile Ala 500 505 510Gln Gly Arg Val Leu Ile Asp Lys Thr Leu Gln Arg Glu Gly Lys Thr 515 520 525Ala Val Lys Leu Glu Asp Leu Ala Thr Arg Leu Gly Phe Lys Thr Pro 530 535 540Glu Asp Leu Phe Ala Ala Val Ala Lys Asp Glu Phe Ser Leu Arg His545 550 555 560Val Glu His Ala Leu Arg His Pro Glu Gly Glu Val Gln Ala Pro Leu 565 570 575Ser Glu Glu Asp Ala Val Thr Lys Lys Ser Arg Ala Thr Ser Val Ala 580 585 590Arg Gly Ala Lys Ser Gly Val Leu Val Val Gly Val Asp Ser Leu Met 595 600 605Thr Gln Met Ser Arg Cys Cys Lys Pro Ala Pro Pro Asp Asp Ile Val 610 615 620Gly Phe Val Thr Arg Gly Arg Gly Val Ser Ile His Arg Arg Ser Cys625 630 635 640His Thr Phe Gln Gln Leu Ala Gly Arg Ala Pro Glu Arg Val Ile Gln 645 650 655Thr Glu Trp Gly Gln Lys Ser His Ala Ala Val Tyr Pro Val Asp Ile 660 665 670His Val Glu Ala Ile Asp Arg Gln Gly Leu Leu Arg Asp Ile Ser Glu 675 680 685Val Leu Ser Arg Glu Lys Ile Asn Val Thr Gly Val Lys Thr Leu Ser 690 695 700Ser Lys Gly Val Ala Arg Met Gln Phe Thr Ala Glu Val Ser Glu Ala705 710 715 720Thr Gln Leu Gln Arg Ala Leu Gln Leu Ile Glu Asp Val Gln Gly Val 725 730 735Leu Gln Ala Lys Arg Lys 740314857DNAC. necator 31atgccgcagg agaacgaaga caaggtcgcg cacctgctgg acgaactggc cagcttcgcg 60cgcgagcggc tgccggcggc gatgttcgcc gtggtcgagc ccttcctgct gcactactac 120gatcaggccg acgccgagga cttgctccag cgcgacgtcg acgatctcta cggcgccgtg 180atggcgcact ggcagaccgc acagaaattc acccccggca atgcgcgcat ccgcgtctac 240aatcccaacc tggaagagca cggctggcac tcggaccaca ccgtggtcga gatcgtcaat 300gacgacatgc ccttcctggt cgactcggtg acgatggaga tcaaccggca gggactggcg 360ctgcactcgg ccatccatcc cgtgttccgg gtctggcgcg acgcgcgcgg cggcatcgag 420cggatcgccc cgggcggcgc cggcgaggcc ggggacagct cgcgcctgga atcctttatc 480catttcgaga tcgaccgcag cggcgaggcc gcgcggctgg aggcgctgcg cagcggcatt 540gcgcaggtgc tggtcgacgt gcgcgcggcg gtggaagact ggtcgaagat gtgcggcatc 600acccaggcca ccatcgccgc catggcgcag gcgcccgacg cggccgcgcc ggagagcgtc 660gaggcgcgcg ccttcctgga ctggatgatg gatgaccact tctccttcct cggccagcgt 720gactaccagc tggtgtcgca ggacggccgc tacttcctgc gcggcgtgcc gggctcgggc 780gcgggcatcc tgcgcgaaag cctgcgcgag cctgatgccg aagatctcac gctgctgccc 840gccgcggcca ccgcgatcat cgaaggggcc tcgccgatct tcctgaccaa ggccaattcg 900cgcgccaccg tgcaccgccc gggctatctc gactatgtcg gcgtcaagct gctcgatgaa 960aaggggcagc tgttcggcga gcggcgcttt gtcggcctgt acacctcgac tgcctatacg 1020gcgccgattg cggaaattcc gctggtgcgg ctcaagtgcg ccaatatcct ggcgcgcgcg 1080ggcttcctgg ccaaggggca cctttacaag tcgctggtga ccatccttga gcaatatccg 1140cgcgatgaac tgttccaggc caccgaggat gaactgttcg acatcaccac cggcatcctg 1200cggctgcagg aacaccagcg cacccggctc tttgtacggc gcgaccgctt cgaccgcttt 1260gtctcgtgcc tggtgttcgt gccgcgcgac aagtacaaca ccgacctgcg ccagaagatc 1320cagaggctgc tgaccgcggc cttccacggc accagctgcg agttcacgcc gctgctgtcc 1380gagtcgccgc tggcgcgcat ccagctgacc gtgcgcggcg agcccggcac catgccgcat 1440gttgatacgc gtgagctgga ggcgcgcatc gtgcacgcca gccgccgctg gcaggacgat 1500ctcgccgaag ccctgcatga aagccacggc gaagagcaag gcaaccggct gctgcagcgc 1560tacggcggct cgttccccgc cggctaccgt gaggactacc cggcccgcac cgcggtgcgc 1620gatatcgagc tgatggagca cgcgctgcgc ggcaacggca tggcgatgaa cctgtaccgg 1680ccgatcgagg ccgcgccggg ggtgttccgc ttcaaggtgt accgtgccgg cgagccgatc 1740gcgctgtcgc acagcctgcc catgctggaa cacctgggcg tgcgcgtgga tgaagagcgc 1800ccctacctga tcgaacccga cagcggcgcg ccggtatggg tgcacgactt cgggctggag 1860attgccgaca gcggcggcgc ggcggaattc gacatcgcgc gcgtcaaggc cttgttcgag 1920gatgcgttcg cgcgcgcctg gcacggcgag atcgagaacg acgacttcaa ccgcctggtg 1980ctgcgcgccg agctggccgc acgcgacgtc accatcctgc

gcgcctatgc ccgctacctg 2040cgccaggtag gctcgacctt cagcgacgcc tatatcgagc gcgcgctgac cggcaacgcg 2100gccattgccg ccatgctggt cggcctgttc gtggcgcgct tcgacacctt cagcgaggtc 2160gccaccgaca ccgcccgcca ggcgcgctgc gacaagctgc tggccgatat cggcgcggcg 2220ctggacaagg tgcccaacct ggacgaggac cgcatcctgc ggcttttcct gggcgtgatc 2280aacgccaccg tgcgcaccaa ctatttccac cgcggcgagg aaggccagcc gcgcccatat 2340gtgtcgttca agttcaatcc cgcgctggtc cccggcctgc ccgagccgcg cccgatgttc 2400gagatctggg tctactcgcc gcgcgttgag ggcgtgcacc tgcgcggcgg gcgcgtggcc 2460cgcggcggac tgcgctggtc agaccggcgc gaggacttcc gcactgaagt gctgggcctg 2520atgaaggcgc agatggtcaa gaacacggtg atcgtgccgg tgggctccaa gggcggcttc 2580gtggtcaagc gcccgccccc gcccaccgac cgcgatgcgt tcctgcagga gggcatcgcc 2640tgctaccaga ccttcctgcg cgggctgctg gacctcaccg acaacctcgt cggcggccag 2700ctggtgccgc cacccgaggt ggtgcgccac gacgacaacg acccctacct ggtggtcgcc 2760gccgacaagg gcaccgcgac gttctccgac ttcgccaacg cgatctcggc cgaatacggc 2820ttctggctgg gcgatgcctt tgcgtccggg ggctcggtcg gctatgacca caagaagatg 2880ggcattaccg cgcgcggtgc gtgggaatcg gtcaagcggc atttccgcga gatgggtgtc 2940gatatccaga ccacggactt caccgtggcc ggcatcggcg acatgtcggg cgatgtgttc 3000ggcaacggca tgctgctgtc gccgcatatc cggctggtgg cggctttcga ccaccggcat 3060atcttcctcg atcccgaccc ggacacgaca aggagcctgc aggaacgcac gcggctgttc 3120ggcctgcccc gctcaagctg ggccgactac gacgccacgc tgatttccgc cggcggcggc 3180atctatccgc gcagcgccaa gaccatcgcg ctgtcgccgc aggtgcaggc ggtgctgggg 3240gtcacggcag ccacgctgtc gccggccgag ctgatccacg ccatcctgat ggcgccggtc 3300gacctgctct acaacggcgg catcggcact tacgtcaagt ccagccagga aacccacctg 3360caggccggcg accgcaccaa cgacgcggtg cgtgtcaacg gcaatgacct gcgctgcaag 3420gtggtcggcg aaggcggcaa cctcggcttc acccagctgg gccgcatcga gttcgcgcgc 3480aagggcgggc gcatcaacac cgacgccatc gacaactcgg ccggggtgga ctgctcggac 3540cacgaggtca atatcaagat cctgctcggg ctggtggttg ccgacggcga aatgaccgag 3600aagcagcgca acaagctgct ggccgagatg accgacgagg tcggcctgct ggtgctgcag 3660gacaactact accagaccca ggcactgtcc gtggccgggc gcagcagccc cgcgctgctc 3720gacggcgagg cgcgcctggt gcgctggctg gagcgcgccg gacggctcaa ccggccgctg 3780gagttcctgc cttcggagga agaaatcgcc gagcgcaagc tggccgatga aggcctggca 3840tcgcccgagc gtgccgtgct gcttgcctac agcaagatgt ggctgtacga cgaactgctg 3900gcctccgacg tgccggaaga cacgctggtc gccggactgc tgtcggacta tttcccggtg 3960ccgctgcgcc agcgttacgc cgacgcgatg cagcgccacc cgctgcggcg cgagatcctg 4020tcgacacacc tgaccaatat gctggtcaac cgtatcggcg ccaccttcgt gcaccggatc 4080atggaagaga ccgacgcacg cccggccgat atcgtgcgcg cctgcctgat cgcacgcgat 4140gtcttcggcc tgaccaccct gtggcaggag atcgatgcgc tggacaaccg tgtggccgat 4200gccgagcagg cgcgcatgtt cggggccgtg gcgctgctgc tggagcgcgc gtgcctgtgg 4260ttcatccgct atctgcgcag cggcagcaag gccgccgagg acctggcgcg ctttgcccag 4320gccgcgcaat ggctggcccc gcaactgccg cggctgctgc cgccggccga tgcgacggca 4380ctgtcagagc gcgcccgggc cttgacggat gcaggggtcg acgaagcgct ggctgtgcgc 4440gtggccggca gcgagatctc cgccgccgcg ctcgatatcg ccgaagtggc cacggcgtgc 4500aaacgcagcc tggacctggt ggcgggggtt tatttcgcgc tggacagcca cctgagcttc 4560agctggctgc gcgagcgcgc gctggcgctg ccatcggaca cgcactggga cctgctggca 4620cgcaccacca cgctggaaga cctggggcgg ctcaagcgcg cactgaccgt cagcgtgctg 4680tcacaggagg gtgaactgga tacgcccgac gccatgatcg atgcctggcg ctccagccgt 4740catggcgcgc tggaacgctt tacgcgcatg ctggccgacc agcgcgcctc gggggctgcg 4800gggctgtcga tgctgtcggt cgcggtgcgc gagatcggca tgctggaacg cgcatag 4857321618PRTC. necator 32Met Pro Gln Glu Asn Glu Asp Lys Val Ala His Leu Leu Asp Glu Leu1 5 10 15Ala Ser Phe Ala Arg Glu Arg Leu Pro Ala Ala Met Phe Ala Val Val 20 25 30Glu Pro Phe Leu Leu His Tyr Tyr Asp Gln Ala Asp Ala Glu Asp Leu 35 40 45Leu Gln Arg Asp Val Asp Asp Leu Tyr Gly Ala Val Met Ala His Trp 50 55 60Gln Thr Ala Gln Lys Phe Thr Pro Gly Asn Ala Arg Ile Arg Val Tyr65 70 75 80Asn Pro Asn Leu Glu Glu His Gly Trp His Ser Asp His Thr Val Val 85 90 95Glu Ile Val Asn Asp Asp Met Pro Phe Leu Val Asp Ser Val Thr Met 100 105 110Glu Ile Asn Arg Gln Gly Leu Ala Leu His Ser Ala Ile His Pro Val 115 120 125Phe Arg Val Trp Arg Asp Ala Arg Gly Gly Ile Glu Arg Ile Ala Pro 130 135 140Gly Gly Ala Gly Glu Ala Gly Asp Ser Ser Arg Leu Glu Ser Phe Ile145 150 155 160His Phe Glu Ile Asp Arg Ser Gly Glu Ala Ala Arg Leu Glu Ala Leu 165 170 175Arg Ser Gly Ile Ala Gln Val Leu Val Asp Val Arg Ala Ala Val Glu 180 185 190Asp Trp Ser Lys Met Cys Gly Ile Thr Gln Ala Thr Ile Ala Ala Met 195 200 205Ala Gln Ala Pro Asp Ala Ala Ala Pro Glu Ser Val Glu Ala Arg Ala 210 215 220Phe Leu Asp Trp Met Met Asp Asp His Phe Ser Phe Leu Gly Gln Arg225 230 235 240Asp Tyr Gln Leu Val Ser Gln Asp Gly Arg Tyr Phe Leu Arg Gly Val 245 250 255Pro Gly Ser Gly Ala Gly Ile Leu Arg Glu Ser Leu Arg Glu Pro Asp 260 265 270Ala Glu Asp Leu Thr Leu Leu Pro Ala Ala Ala Thr Ala Ile Ile Glu 275 280 285Gly Ala Ser Pro Ile Phe Leu Thr Lys Ala Asn Ser Arg Ala Thr Val 290 295 300His Arg Pro Gly Tyr Leu Asp Tyr Val Gly Val Lys Leu Leu Asp Glu305 310 315 320Lys Gly Gln Leu Phe Gly Glu Arg Arg Phe Val Gly Leu Tyr Thr Ser 325 330 335Thr Ala Tyr Thr Ala Pro Ile Ala Glu Ile Pro Leu Val Arg Leu Lys 340 345 350Cys Ala Asn Ile Leu Ala Arg Ala Gly Phe Leu Ala Lys Gly His Leu 355 360 365Tyr Lys Ser Leu Val Thr Ile Leu Glu Gln Tyr Pro Arg Asp Glu Leu 370 375 380Phe Gln Ala Thr Glu Asp Glu Leu Phe Asp Ile Thr Thr Gly Ile Leu385 390 395 400Arg Leu Gln Glu His Gln Arg Thr Arg Leu Phe Val Arg Arg Asp Arg 405 410 415Phe Asp Arg Phe Val Ser Cys Leu Val Phe Val Pro Arg Asp Lys Tyr 420 425 430Asn Thr Asp Leu Arg Gln Lys Ile Gln Arg Leu Leu Thr Ala Ala Phe 435 440 445His Gly Thr Ser Cys Glu Phe Thr Pro Leu Leu Ser Glu Ser Pro Leu 450 455 460Ala Arg Ile Gln Leu Thr Val Arg Gly Glu Pro Gly Thr Met Pro His465 470 475 480Val Asp Thr Arg Glu Leu Glu Ala Arg Ile Val His Ala Ser Arg Arg 485 490 495Trp Gln Asp Asp Leu Ala Glu Ala Leu His Glu Ser His Gly Glu Glu 500 505 510Gln Gly Asn Arg Leu Leu Gln Arg Tyr Gly Gly Ser Phe Pro Ala Gly 515 520 525Tyr Arg Glu Asp Tyr Pro Ala Arg Thr Ala Val Arg Asp Ile Glu Leu 530 535 540Met Glu His Ala Leu Arg Gly Asn Gly Met Ala Met Asn Leu Tyr Arg545 550 555 560Pro Ile Glu Ala Ala Pro Gly Val Phe Arg Phe Lys Val Tyr Arg Ala 565 570 575Gly Glu Pro Ile Ala Leu Ser His Ser Leu Pro Met Leu Glu His Leu 580 585 590Gly Val Arg Val Asp Glu Glu Arg Pro Tyr Leu Ile Glu Pro Asp Ser 595 600 605Gly Ala Pro Val Trp Val His Asp Phe Gly Leu Glu Ile Ala Asp Ser 610 615 620Gly Gly Ala Ala Glu Phe Asp Ile Ala Arg Val Lys Ala Leu Phe Glu625 630 635 640Asp Ala Phe Ala Arg Ala Trp His Gly Glu Ile Glu Asn Asp Asp Phe 645 650 655Asn Arg Leu Val Leu Arg Ala Glu Leu Ala Ala Arg Asp Val Thr Ile 660 665 670Leu Arg Ala Tyr Ala Arg Tyr Leu Arg Gln Val Gly Ser Thr Phe Ser 675 680 685Asp Ala Tyr Ile Glu Arg Ala Leu Thr Gly Asn Ala Ala Ile Ala Ala 690 695 700Met Leu Val Gly Leu Phe Val Ala Arg Phe Asp Thr Phe Ser Glu Val705 710 715 720Ala Thr Asp Thr Ala Arg Gln Ala Arg Cys Asp Lys Leu Leu Ala Asp 725 730 735Ile Gly Ala Ala Leu Asp Lys Val Pro Asn Leu Asp Glu Asp Arg Ile 740 745 750Leu Arg Leu Phe Leu Gly Val Ile Asn Ala Thr Val Arg Thr Asn Tyr 755 760 765Phe His Arg Gly Glu Glu Gly Gln Pro Arg Pro Tyr Val Ser Phe Lys 770 775 780Phe Asn Pro Ala Leu Val Pro Gly Leu Pro Glu Pro Arg Pro Met Phe785 790 795 800Glu Ile Trp Val Tyr Ser Pro Arg Val Glu Gly Val His Leu Arg Gly 805 810 815Gly Arg Val Ala Arg Gly Gly Leu Arg Trp Ser Asp Arg Arg Glu Asp 820 825 830Phe Arg Thr Glu Val Leu Gly Leu Met Lys Ala Gln Met Val Lys Asn 835 840 845Thr Val Ile Val Pro Val Gly Ser Lys Gly Gly Phe Val Val Lys Arg 850 855 860Pro Pro Pro Pro Thr Asp Arg Asp Ala Phe Leu Gln Glu Gly Ile Ala865 870 875 880Cys Tyr Gln Thr Phe Leu Arg Gly Leu Leu Asp Leu Thr Asp Asn Leu 885 890 895Val Gly Gly Gln Leu Val Pro Pro Pro Glu Val Val Arg His Asp Asp 900 905 910Asn Asp Pro Tyr Leu Val Val Ala Ala Asp Lys Gly Thr Ala Thr Phe 915 920 925Ser Asp Phe Ala Asn Ala Ile Ser Ala Glu Tyr Gly Phe Trp Leu Gly 930 935 940Asp Ala Phe Ala Ser Gly Gly Ser Val Gly Tyr Asp His Lys Lys Met945 950 955 960Gly Ile Thr Ala Arg Gly Ala Trp Glu Ser Val Lys Arg His Phe Arg 965 970 975Glu Met Gly Val Asp Ile Gln Thr Thr Asp Phe Thr Val Ala Gly Ile 980 985 990Gly Asp Met Ser Gly Asp Val Phe Gly Asn Gly Met Leu Leu Ser Pro 995 1000 1005His Ile Arg Leu Val Ala Ala Phe Asp His Arg His Ile Phe Leu 1010 1015 1020Asp Pro Asp Pro Asp Thr Thr Arg Ser Leu Gln Glu Arg Thr Arg 1025 1030 1035Leu Phe Gly Leu Pro Arg Ser Ser Trp Ala Asp Tyr Asp Ala Thr 1040 1045 1050Leu Ile Ser Ala Gly Gly Gly Ile Tyr Pro Arg Ser Ala Lys Thr 1055 1060 1065Ile Ala Leu Ser Pro Gln Val Gln Ala Val Leu Gly Val Thr Ala 1070 1075 1080Ala Thr Leu Ser Pro Ala Glu Leu Ile His Ala Ile Leu Met Ala 1085 1090 1095Pro Val Asp Leu Leu Tyr Asn Gly Gly Ile Gly Thr Tyr Val Lys 1100 1105 1110Ser Ser Gln Glu Thr His Leu Gln Ala Gly Asp Arg Thr Asn Asp 1115 1120 1125Ala Val Arg Val Asn Gly Asn Asp Leu Arg Cys Lys Val Val Gly 1130 1135 1140Glu Gly Gly Asn Leu Gly Phe Thr Gln Leu Gly Arg Ile Glu Phe 1145 1150 1155Ala Arg Lys Gly Gly Arg Ile Asn Thr Asp Ala Ile Asp Asn Ser 1160 1165 1170Ala Gly Val Asp Cys Ser Asp His Glu Val Asn Ile Lys Ile Leu 1175 1180 1185Leu Gly Leu Val Val Ala Asp Gly Glu Met Thr Glu Lys Gln Arg 1190 1195 1200Asn Lys Leu Leu Ala Glu Met Thr Asp Glu Val Gly Leu Leu Val 1205 1210 1215Leu Gln Asp Asn Tyr Tyr Gln Thr Gln Ala Leu Ser Val Ala Gly 1220 1225 1230Arg Ser Ser Pro Ala Leu Leu Asp Gly Glu Ala Arg Leu Val Arg 1235 1240 1245Trp Leu Glu Arg Ala Gly Arg Leu Asn Arg Pro Leu Glu Phe Leu 1250 1255 1260Pro Ser Glu Glu Glu Ile Ala Glu Arg Lys Leu Ala Asp Glu Gly 1265 1270 1275Leu Ala Ser Pro Glu Arg Ala Val Leu Leu Ala Tyr Ser Lys Met 1280 1285 1290Trp Leu Tyr Asp Glu Leu Leu Ala Ser Asp Val Pro Glu Asp Thr 1295 1300 1305Leu Val Ala Gly Leu Leu Ser Asp Tyr Phe Pro Val Pro Leu Arg 1310 1315 1320Gln Arg Tyr Ala Asp Ala Met Gln Arg His Pro Leu Arg Arg Glu 1325 1330 1335Ile Leu Ser Thr His Leu Thr Asn Met Leu Val Asn Arg Ile Gly 1340 1345 1350Ala Thr Phe Val His Arg Ile Met Glu Glu Thr Asp Ala Arg Pro 1355 1360 1365Ala Asp Ile Val Arg Ala Cys Leu Ile Ala Arg Asp Val Phe Gly 1370 1375 1380Leu Thr Thr Leu Trp Gln Glu Ile Asp Ala Leu Asp Asn Arg Val 1385 1390 1395Ala Asp Ala Glu Gln Ala Arg Met Phe Gly Ala Val Ala Leu Leu 1400 1405 1410Leu Glu Arg Ala Cys Leu Trp Phe Ile Arg Tyr Leu Arg Ser Gly 1415 1420 1425Ser Lys Ala Ala Glu Asp Leu Ala Arg Phe Ala Gln Ala Ala Gln 1430 1435 1440Trp Leu Ala Pro Gln Leu Pro Arg Leu Leu Pro Pro Ala Asp Ala 1445 1450 1455Thr Ala Leu Ser Glu Arg Ala Arg Ala Leu Thr Asp Ala Gly Val 1460 1465 1470Asp Glu Ala Leu Ala Val Arg Val Ala Gly Ser Glu Ile Ser Ala 1475 1480 1485Ala Ala Leu Asp Ile Ala Glu Val Ala Thr Ala Cys Lys Arg Ser 1490 1495 1500Leu Asp Leu Val Ala Gly Val Tyr Phe Ala Leu Asp Ser His Leu 1505 1510 1515Ser Phe Ser Trp Leu Arg Glu Arg Ala Leu Ala Leu Pro Ser Asp 1520 1525 1530Thr His Trp Asp Leu Leu Ala Arg Thr Thr Thr Leu Glu Asp Leu 1535 1540 1545Gly Arg Leu Lys Arg Ala Leu Thr Val Ser Val Leu Ser Gln Glu 1550 1555 1560Gly Glu Leu Asp Thr Pro Asp Ala Met Ile Asp Ala Trp Arg Ser 1565 1570 1575Ser Arg His Gly Ala Leu Glu Arg Phe Thr Arg Met Leu Ala Asp 1580 1585 1590Gln Arg Ala Ser Gly Ala Ala Gly Leu Ser Met Leu Ser Val Ala 1595 1600 1605Val Arg Glu Ile Gly Met Leu Glu Arg Ala 1610 161533633DNAC. necator 33atgaccgcaa cgccaaaccc cacgccccac cgcggcgaga ccgtgttcat cgtcgacgac 60gatgaagcca tgcgcgactc gctgacctgg ctgctggagg gcaatggcta ccaggtgcgc 120agcttcacca gcgccgagca gttcctcgcc gcctacgatg ccagccaggt gtcgtgcctg 180atcctcgacg tgcgcatgcc cggcatgagc ggcccggaac tgcaggagcg catgctggcc 240gagcagatcg acattcccat cgtctttatc accggccacg gcgacgtgcc gatggcggta 300tcgacgatga agcgcggcgc catcgacttc atcgaaaagc ccttcgatga gtccgagctg 360cgcgcactgg tcgagcgcat gctgaccaag gcccgcaccg accattccgc cgcacgcgag 420cagcgcgccg ccaaggacct gctgggcaag ctgaccacgc gcgagcagca ggtgctcgag 480cgcatcgttg ccggccgcct gaacaagcag atcgccgacg acctgggcat ttccatcaag 540accgtggagg cgcaccgcgc caacatcatg gaaaagctca acgtcaacac cgtcgccgac 600ctgctgcgcc tggcgctgtc acgcaacagc tga 63334210PRTC. necator 34Met Thr Ala Thr Pro Asn Pro Thr Pro His Arg Gly Glu Thr Val Phe1 5 10 15Ile Val Asp Asp Asp Glu Ala Met Arg Asp Ser Leu Thr Trp Leu Leu 20 25 30Glu Gly Asn Gly Tyr Gln Val Arg Ser Phe Thr Ser Ala Glu Gln Phe 35 40 45Leu Ala Ala Tyr Asp Ala Ser Gln Val Ser Cys Leu Ile Leu Asp Val 50 55 60Arg Met Pro Gly Met Ser Gly Pro Glu Leu Gln Glu Arg Met Leu Ala65 70 75 80Glu Gln Ile Asp Ile Pro Ile Val Phe Ile Thr Gly His Gly Asp Val 85 90 95Pro Met Ala Val Ser Thr Met Lys Arg Gly Ala Ile Asp Phe Ile Glu 100 105 110Lys Pro Phe Asp Glu Ser Glu Leu Arg Ala Leu Val Glu Arg Met Leu 115 120 125Thr Lys Ala Arg Thr Asp His Ser Ala Ala Arg Glu Gln Arg Ala Ala 130 135 140Lys Asp Leu Leu Gly Lys Leu Thr Thr Arg Glu Gln Gln Val Leu Glu145 150 155 160Arg Ile Val Ala Gly Arg Leu Asn Lys Gln Ile Ala Asp Asp Leu Gly 165 170 175Ile Ser Ile Lys Thr Val Glu Ala His Arg Ala Asn Ile Met Glu Lys 180 185 190Leu Asn Val Asn Thr Val Ala Asp Leu Leu Arg Leu Ala Leu Ser Arg 195 200 205Asn Ser 210352586DNAC. necator 35atgccgtttt tcgaccgact cctttccagc ctgcgtgcag ccatgcttcc tcccgacgcg 60tcggggtcag ccaaggcggc cggaggcgat cccgcgcacg ccccttcgcc ggctgccgga 120ctggggccga tcgaggcgct gggccccgac gacggcgccc ccgtggccgc accgcgcggg 180ctgtggtggc tgcgctggcg caacctggtg gccaccagct ggttcatgtt catcccgctg 240gttgccatcg tgctgttcac ggtggccatg ggcgtgatcc tgtggtccct gcacgagacc

300gagcgccagc agcagcgcga cgccctgtac cgcgacgccg cctgggcgca gcagcgcgtg 360cgcctgtccc tgctgagcaa ccaggaccag ctggcctcgc tggcgcgcga catcgccgcc 420gcgcagcttg agcaaggcgc ctaccgcacc gcggcgcagg aaatcctgcg cgaaaacccc 480gagatcgttt tcatcaactg gctcgatgcc accaagcgcg gccgctggtc gttgccatcg 540acctcggaat tcgccagccg cctgcgcgag aaccaggacc agccgctcga accggaagtg 600ctcgacacct tcgacgccgc gcgcgagacc cagcgagtgg tctattcgcg cccgctggtc 660aacgagcgcg gcgacagctt catgctgatg gaagtgccga tcgtgcgcga caacgagttc 720ctcggcacgc tcggcgcgct gtactcgatc aacggcatcc tgacgcacct gctgccgcct 780gagctgaccg agcgctaccg cttctcgctg atcgacaaga acaaccagac ccgcgccagc 840acctcgttgc ggccggtgcc gggcaacgcg ctgtcgtacg aggtgctgct ggatccgccg 900ggtcactccc tgtcgctgcg cgccgatgcc tacccgccgg cgtcgaacct gcccaacaac 960atgctgctgt ggctggtggt ggggctgtcg tgcttcctgc tgtggagcct gtggagcatg 1020tggcgccaca ccagccgccg ctccgaggcg cagcgcgcgc tgctggccga gacctcgttc 1080cggcgcgcga tggaaaactc gatgctgatc ggcctgcgcg cgctcgacct gaacggccgc 1140atcacctatg tcaacccggc cttctgccgc atgactggct ggcaggagaa cgacctggtc 1200gggcgcctgc cgcccttccc ctactggccg cccaacgacc agcaggagat gcagaagcag 1260atcgacctga cgctgcaggg caagtcgccc gccggcggct acgagatgcg cgtgatgcgc 1320cgcgacggca gcagcttcta cgcgcgcatg tacgtgtcgc cgctggtgga cagccgcggc 1380cgccatactg gctggatgag ctcgatgacg gacatcaccg agcccaagcg cgcgcgcgag 1440gaactggccg cggcgcatga ccgcttcacc acggtgctgg aaagcctgga tgccgcggtg 1500tcggtgctgg ccaccgacaa ggcggagctg ctctttgcca accgctatta ccgccagctg 1560ttcggctggg aggccgaggg ccacctcaag ctggccggcg acgacctcga caaggaccag 1620gtctccagcg acaacaccga ctatgtcgac gcctatgccg gcctgccggc gtccgagctg 1680atgccgtacg catcggacgc gcgcgaagtg ttcgtgccgg acatgcagaa atggttcgaa 1740gtgcgccgcc gctatatcca gtgggtcgac ggccacctgg cgcagatgca gatcgccacc 1800gatatcacgg tgcgcaaggc cgccgaggaa atggcgcgcc agcatgaaga gcgcctgcag 1860ttcaccagcc gcctgaccac catgggcgag atggcctcgt cgctggcgca tgaactgaac 1920cagccgctgg cggccatcaa caactactgc atgggcgcgg tggcgcggct gcactcgggc 1980cgcagcacgc cggaggacct gatcccggtg ctggagaaga cctcggccca ggcggtgcgc 2040gccggcacca tcatcagccg catccgcggc ttcgtgaagc gcagccagcc gcagcggcgc 2100gaggccgcgc tgcacgacat cgtcgccgac gcggtaggcc tggccgacct ggaggccacg 2160cgccgccgcg tcaccatcct gacccgcctg ccgaccccgc cgctgacggt ctatgtcgac 2220ccggtgctga tcgagcaggt gctggtgaac ctgctcaaga acgcggtcga ggccatggcc 2280ggcctgcccg ccctgcacgc cggcggcgtg gtgcgcctgc acgcgcgggt ggagccgggc 2340gagattggcg acagcgtcca tatcgacgtg atcgaccagg gcccgggcgt ggacgaagcc 2400accaaggagc gcctgttcga gcccttcttc agcaccaagt ccgacggcat gggcatgggg 2460ctgaacatct gccgctcgat catcgagtcg caccagggcc gcctgtgggt ggagaacaat 2520gccgacggca tcggctgtac atttaaaatc atgctgccgc tgcaatcggc gctggccgag 2580cattaa 258636861PRTC. necator 36Met Pro Phe Phe Asp Arg Leu Leu Ser Ser Leu Arg Ala Ala Met Leu1 5 10 15Pro Pro Asp Ala Ser Gly Ser Ala Lys Ala Ala Gly Gly Asp Pro Ala 20 25 30His Ala Pro Ser Pro Ala Ala Gly Leu Gly Pro Ile Glu Ala Leu Gly 35 40 45Pro Asp Asp Gly Ala Pro Val Ala Ala Pro Arg Gly Leu Trp Trp Leu 50 55 60Arg Trp Arg Asn Leu Val Ala Thr Ser Trp Phe Met Phe Ile Pro Leu65 70 75 80Val Ala Ile Val Leu Phe Thr Val Ala Met Gly Val Ile Leu Trp Ser 85 90 95Leu His Glu Thr Glu Arg Gln Gln Gln Arg Asp Ala Leu Tyr Arg Asp 100 105 110Ala Ala Trp Ala Gln Gln Arg Val Arg Leu Ser Leu Leu Ser Asn Gln 115 120 125Asp Gln Leu Ala Ser Leu Ala Arg Asp Ile Ala Ala Ala Gln Leu Glu 130 135 140Gln Gly Ala Tyr Arg Thr Ala Ala Gln Glu Ile Leu Arg Glu Asn Pro145 150 155 160Glu Ile Val Phe Ile Asn Trp Leu Asp Ala Thr Lys Arg Gly Arg Trp 165 170 175Ser Leu Pro Ser Thr Ser Glu Phe Ala Ser Arg Leu Arg Glu Asn Gln 180 185 190Asp Gln Pro Leu Glu Pro Glu Val Leu Asp Thr Phe Asp Ala Ala Arg 195 200 205Glu Thr Gln Arg Val Val Tyr Ser Arg Pro Leu Val Asn Glu Arg Gly 210 215 220Asp Ser Phe Met Leu Met Glu Val Pro Ile Val Arg Asp Asn Glu Phe225 230 235 240Leu Gly Thr Leu Gly Ala Leu Tyr Ser Ile Asn Gly Ile Leu Thr His 245 250 255Leu Leu Pro Pro Glu Leu Thr Glu Arg Tyr Arg Phe Ser Leu Ile Asp 260 265 270Lys Asn Asn Gln Thr Arg Ala Ser Thr Ser Leu Arg Pro Val Pro Gly 275 280 285Asn Ala Leu Ser Tyr Glu Val Leu Leu Asp Pro Pro Gly His Ser Leu 290 295 300Ser Leu Arg Ala Asp Ala Tyr Pro Pro Ala Ser Asn Leu Pro Asn Asn305 310 315 320Met Leu Leu Trp Leu Val Val Gly Leu Ser Cys Phe Leu Leu Trp Ser 325 330 335Leu Trp Ser Met Trp Arg His Thr Ser Arg Arg Ser Glu Ala Gln Arg 340 345 350Ala Leu Leu Ala Glu Thr Ser Phe Arg Arg Ala Met Glu Asn Ser Met 355 360 365Leu Ile Gly Leu Arg Ala Leu Asp Leu Asn Gly Arg Ile Thr Tyr Val 370 375 380Asn Pro Ala Phe Cys Arg Met Thr Gly Trp Gln Glu Asn Asp Leu Val385 390 395 400Gly Arg Leu Pro Pro Phe Pro Tyr Trp Pro Pro Asn Asp Gln Gln Glu 405 410 415Met Gln Lys Gln Ile Asp Leu Thr Leu Gln Gly Lys Ser Pro Ala Gly 420 425 430Gly Tyr Glu Met Arg Val Met Arg Arg Asp Gly Ser Ser Phe Tyr Ala 435 440 445Arg Met Tyr Val Ser Pro Leu Val Asp Ser Arg Gly Arg His Thr Gly 450 455 460Trp Met Ser Ser Met Thr Asp Ile Thr Glu Pro Lys Arg Ala Arg Glu465 470 475 480Glu Leu Ala Ala Ala His Asp Arg Phe Thr Thr Val Leu Glu Ser Leu 485 490 495Asp Ala Ala Val Ser Val Leu Ala Thr Asp Lys Ala Glu Leu Leu Phe 500 505 510Ala Asn Arg Tyr Tyr Arg Gln Leu Phe Gly Trp Glu Ala Glu Gly His 515 520 525Leu Lys Leu Ala Gly Asp Asp Leu Asp Lys Asp Gln Val Ser Ser Asp 530 535 540Asn Thr Asp Tyr Val Asp Ala Tyr Ala Gly Leu Pro Ala Ser Glu Leu545 550 555 560Met Pro Tyr Ala Ser Asp Ala Arg Glu Val Phe Val Pro Asp Met Gln 565 570 575Lys Trp Phe Glu Val Arg Arg Arg Tyr Ile Gln Trp Val Asp Gly His 580 585 590Leu Ala Gln Met Gln Ile Ala Thr Asp Ile Thr Val Arg Lys Ala Ala 595 600 605Glu Glu Met Ala Arg Gln His Glu Glu Arg Leu Gln Phe Thr Ser Arg 610 615 620Leu Thr Thr Met Gly Glu Met Ala Ser Ser Leu Ala His Glu Leu Asn625 630 635 640Gln Pro Leu Ala Ala Ile Asn Asn Tyr Cys Met Gly Ala Val Ala Arg 645 650 655Leu His Ser Gly Arg Ser Thr Pro Glu Asp Leu Ile Pro Val Leu Glu 660 665 670Lys Thr Ser Ala Gln Ala Val Arg Ala Gly Thr Ile Ile Ser Arg Ile 675 680 685Arg Gly Phe Val Lys Arg Ser Gln Pro Gln Arg Arg Glu Ala Ala Leu 690 695 700His Asp Ile Val Ala Asp Ala Val Gly Leu Ala Asp Leu Glu Ala Thr705 710 715 720Arg Arg Arg Val Thr Ile Leu Thr Arg Leu Pro Thr Pro Pro Leu Thr 725 730 735Val Tyr Val Asp Pro Val Leu Ile Glu Gln Val Leu Val Asn Leu Leu 740 745 750Lys Asn Ala Val Glu Ala Met Ala Gly Leu Pro Ala Leu His Ala Gly 755 760 765Gly Val Val Arg Leu His Ala Arg Val Glu Pro Gly Glu Ile Gly Asp 770 775 780Ser Val His Ile Asp Val Ile Asp Gln Gly Pro Gly Val Asp Glu Ala785 790 795 800Thr Lys Glu Arg Leu Phe Glu Pro Phe Phe Ser Thr Lys Ser Asp Gly 805 810 815Met Gly Met Gly Leu Asn Ile Cys Arg Ser Ile Ile Glu Ser His Gln 820 825 830Gly Arg Leu Trp Val Glu Asn Asn Ala Asp Gly Ile Gly Cys Thr Phe 835 840 845Lys Ile Met Leu Pro Leu Gln Ser Ala Leu Ala Glu His 850 855 860372583DNAC. necator 37atggacacca cgccggaact gctgcttgcc gcgcgcgtgc gcgaccagct caaagccgac 60aagcaggcac tgtttgccga cttcaacatc agcgccaatg tcggcacgct gatcacgcgg 120ctgcgccgcg ccgtcgacgc cgggctggtg gaagcctggc gcggcctggg gatgcccgcg 180ggcgcggcgc tggtggcggt gggcggctat ggccgcggcg agctgctgcc ttactccgac 240gtcgacgtgc tgctgttgct gcccgccgag ccagaccagg acaccaccgg gcgcctggag 300cgctttatcg gcctgtgctg ggacctcggg ctggagatcg gctcttcggt gcgcaccgtg 360gacgattgca tccgcgagtc gcgccaggac gtcaccatcc agacctcgct gctggaagcg 420cggctgctga ccggcagccg caagctgttc gagtcgatgc gtacgcgcta cctcgccgac 480ctggacccgg ccgcgttctt ccaggccaag ctgctggaaa tgcgccagcg ccacgccaag 540taccaggaca cgccctactc gctcgagccc aactgcaagg aaagccccgg cggcctgcgc 600gatctgcagg tgatcctgtg gatgaccaag gcggcgggcc tgggcgacag ctggaaagaa 660cttttcgagc gcggcctgct gacgcagcgc gaagcgcagg agctcgcgcg caacgagcgc 720ctgctcaaga ccatccgcgc gcgcctgcac ctggtggccg gccggcgcca ggacgtgctg 780gtattcgacc tgcagaccgc gctggcggag tccttcggct atcgccagac caccagcaag 840cgcgccagcg aacaactgat gcgccgctac tactgggcgg ccaaggcagt cacgcagctc 900aacagcgtgc tgctgctgaa catcgaggcg atgctgttcc cgagcgagtc tcaggtgacg 960cgcgtgctca acgagcgctt tgtcgagcgc cagggcatgc tggaaatcac cagcgacgac 1020atctatgaac gcgacccgca cgcgatcctg gaaaccttcc tgctgtacca gcgcacgccc 1080ggcgtgaaag gcctgtcgcc gcgcacgctg cgcgggctgt acaacgcgcg caccgtgatg 1140aatgcgggct ggcgcaacga tccggaaaac cgccgcctgt tcctggccat catgcaggag 1200ccgcagggca tcacccacgc gctgcgcctg atgaaccaga ccagcgtgct gggccgctac 1260ctgatcaact tccggcgcat cgtcggccag atgcagcacg acctgttcca cgtctacacc 1320gtggaccagc acatcctgat ggtggtgcgc aacatgcgcc gcttcgccat cgtcgagcac 1380acccacgagt tcccgttctg cagccagctg atggccagct tcgacaagcc gtgggtgctg 1440tgggtggcag cgctgttcca cgacatcgcc aagggccgcg gcggcgacca ctcgaagctt 1500ggcaccgtgg atgcgcgccg cttctgcaag cagcacggca ttgcgcgcga agacgccgac 1560ctggtctgct ggctggtcga gcaccacctg accatgagcc acgtggcgca gaagcaggac 1620ctgaccgatc ccgatgtgat ccacgccttt gcccgcgtgg tcggcagcga acgctacctg 1680accgcgctct acctgctgac cgtggccgac atccgcggca ccagtcccaa ggtatggaac 1740gcgtggaagg gcaagctgct ggaagacctg taccacatca cgctgcgcgt gctcggcggc 1800gcgcgcgtgg attcgcattc gctgtggtcg cagcgcaagg aagacaccat ctccgagctg 1860cgcctgaagg ccttcgaccc ggcgctgggc aagtccctgt gggcgcagct cgacgtggct 1920ttcttcctgc gccacgattc gcacgatatc gcctggctca cgcgccacct gtacaacaag 1980gtggacagcc ccacgccggt ggtcaaggca cgcgtctccc ccgccggcga aggcctgcag 2040gtggcggtct acgtcaagga ccagcccgac ctgttcgcgc gcatctgcgg ctacttcgag 2100cgcaaggcat tctcgatcca ggacgccaag atccacacca cgcgccacgg ctacgcgctg 2160gacacgttcc aggtcaccga ccccggcatg gccggcgacg gcggcagcta ccgcgacatc 2220atcgcgctgg tcgagcacga actgtgcgag cggctgcgcc tgcaaggcgc actgcccgaa 2280cccacgcagg ggcggctgtc gcgccagtcg cgcagcttcc cgatcaagcc gcgcgtggac 2340ctgcgccccg acgagcgcgg ccagtattac ctgctgtcgc tgtccgccaa cgaccgcacc 2400ggcctgctgt acgccatcgc ccgcgtactg gcacggcatc gcgtatccgt ccacacggca 2460cgcatcaaca ccctgggcga acgcgtcgaa gacgtgttcc tggtcgacgg cagccgcctg 2520gctgccgaca accgattgca gattcagctt gaacaggact tgctcgccgc cctcgccatc 2580tga 258338860PRTC. necator 38Met Asp Thr Thr Pro Glu Leu Leu Leu Ala Ala Arg Val Arg Asp Gln1 5 10 15Leu Lys Ala Asp Lys Gln Ala Leu Phe Ala Asp Phe Asn Ile Ser Ala 20 25 30Asn Val Gly Thr Leu Ile Thr Arg Leu Arg Arg Ala Val Asp Ala Gly 35 40 45Leu Val Glu Ala Trp Arg Gly Leu Gly Met Pro Ala Gly Ala Ala Leu 50 55 60Val Ala Val Gly Gly Tyr Gly Arg Gly Glu Leu Leu Pro Tyr Ser Asp65 70 75 80Val Asp Val Leu Leu Leu Leu Pro Ala Glu Pro Asp Gln Asp Thr Thr 85 90 95Gly Arg Leu Glu Arg Phe Ile Gly Leu Cys Trp Asp Leu Gly Leu Glu 100 105 110Ile Gly Ser Ser Val Arg Thr Val Asp Asp Cys Ile Arg Glu Ser Arg 115 120 125Gln Asp Val Thr Ile Gln Thr Ser Leu Leu Glu Ala Arg Leu Leu Thr 130 135 140Gly Ser Arg Lys Leu Phe Glu Ser Met Arg Thr Arg Tyr Leu Ala Asp145 150 155 160Leu Asp Pro Ala Ala Phe Phe Gln Ala Lys Leu Leu Glu Met Arg Gln 165 170 175Arg His Ala Lys Tyr Gln Asp Thr Pro Tyr Ser Leu Glu Pro Asn Cys 180 185 190Lys Glu Ser Pro Gly Gly Leu Arg Asp Leu Gln Val Ile Leu Trp Met 195 200 205Thr Lys Ala Ala Gly Leu Gly Asp Ser Trp Lys Glu Leu Phe Glu Arg 210 215 220Gly Leu Leu Thr Gln Arg Glu Ala Gln Glu Leu Ala Arg Asn Glu Arg225 230 235 240Leu Leu Lys Thr Ile Arg Ala Arg Leu His Leu Val Ala Gly Arg Arg 245 250 255Gln Asp Val Leu Val Phe Asp Leu Gln Thr Ala Leu Ala Glu Ser Phe 260 265 270Gly Tyr Arg Gln Thr Thr Ser Lys Arg Ala Ser Glu Gln Leu Met Arg 275 280 285Arg Tyr Tyr Trp Ala Ala Lys Ala Val Thr Gln Leu Asn Ser Val Leu 290 295 300Leu Leu Asn Ile Glu Ala Met Leu Phe Pro Ser Glu Ser Gln Val Thr305 310 315 320Arg Val Leu Asn Glu Arg Phe Val Glu Arg Gln Gly Met Leu Glu Ile 325 330 335Thr Ser Asp Asp Ile Tyr Glu Arg Asp Pro His Ala Ile Leu Glu Thr 340 345 350Phe Leu Leu Tyr Gln Arg Thr Pro Gly Val Lys Gly Leu Ser Pro Arg 355 360 365Thr Leu Arg Gly Leu Tyr Asn Ala Arg Thr Val Met Asn Ala Gly Trp 370 375 380Arg Asn Asp Pro Glu Asn Arg Arg Leu Phe Leu Ala Ile Met Gln Glu385 390 395 400Pro Gln Gly Ile Thr His Ala Leu Arg Leu Met Asn Gln Thr Ser Val 405 410 415Leu Gly Arg Tyr Leu Ile Asn Phe Arg Arg Ile Val Gly Gln Met Gln 420 425 430His Asp Leu Phe His Val Tyr Thr Val Asp Gln His Ile Leu Met Val 435 440 445Val Arg Asn Met Arg Arg Phe Ala Ile Val Glu His Thr His Glu Phe 450 455 460Pro Phe Cys Ser Gln Leu Met Ala Ser Phe Asp Lys Pro Trp Val Leu465 470 475 480Trp Val Ala Ala Leu Phe His Asp Ile Ala Lys Gly Arg Gly Gly Asp 485 490 495His Ser Lys Leu Gly Thr Val Asp Ala Arg Arg Phe Cys Lys Gln His 500 505 510Gly Ile Ala Arg Glu Asp Ala Asp Leu Val Cys Trp Leu Val Glu His 515 520 525His Leu Thr Met Ser His Val Ala Gln Lys Gln Asp Leu Thr Asp Pro 530 535 540Asp Val Ile His Ala Phe Ala Arg Val Val Gly Ser Glu Arg Tyr Leu545 550 555 560Thr Ala Leu Tyr Leu Leu Thr Val Ala Asp Ile Arg Gly Thr Ser Pro 565 570 575Lys Val Trp Asn Ala Trp Lys Gly Lys Leu Leu Glu Asp Leu Tyr His 580 585 590Ile Thr Leu Arg Val Leu Gly Gly Ala Arg Val Asp Ser His Ser Leu 595 600 605Trp Ser Gln Arg Lys Glu Asp Thr Ile Ser Glu Leu Arg Leu Lys Ala 610 615 620Phe Asp Pro Ala Leu Gly Lys Ser Leu Trp Ala Gln Leu Asp Val Ala625 630 635 640Phe Phe Leu Arg His Asp Ser His Asp Ile Ala Trp Leu Thr Arg His 645 650 655Leu Tyr Asn Lys Val Asp Ser Pro Thr Pro Val Val Lys Ala Arg Val 660 665 670Ser Pro Ala Gly Glu Gly Leu Gln Val Ala Val Tyr Val Lys Asp Gln 675 680 685Pro Asp Leu Phe Ala Arg Ile Cys Gly Tyr Phe Glu Arg Lys Ala Phe 690 695 700Ser Ile Gln Asp Ala Lys Ile His Thr Thr Arg His Gly Tyr Ala Leu705 710 715 720Asp Thr Phe Gln Val Thr Asp Pro Gly Met Ala Gly Asp Gly Gly Ser 725 730 735Tyr Arg Asp Ile Ile Ala Leu Val Glu His Glu Leu Cys Glu Arg Leu 740 745 750Arg Leu Gln Gly Ala Leu Pro Glu Pro Thr Gln Gly Arg Leu Ser Arg 755 760 765Gln Ser Arg Ser Phe Pro Ile Lys Pro Arg Val Asp Leu Arg Pro Asp 770 775 780Glu Arg Gly Gln Tyr

Tyr Leu Leu Ser Leu Ser Ala Asn Asp Arg Thr785 790 795 800Gly Leu Leu Tyr Ala Ile Ala Arg Val Leu Ala Arg His Arg Val Ser 805 810 815Val His Thr Ala Arg Ile Asn Thr Leu Gly Glu Arg Val Glu Asp Val 820 825 830Phe Leu Val Asp Gly Ser Arg Leu Ala Ala Asp Asn Arg Leu Gln Ile 835 840 845Gln Leu Glu Gln Asp Leu Leu Ala Ala Leu Ala Ile 850 855 860391548DNAC. necator 39atgaagccga tctggatagt cgacgacgat caatcaatcc gctgggtcct ggaaaaggcc 60ctggcccgtg aaagcctgct ctcgcgcagc ttcaccaatg tgcgggatgc gctggccgcg 120ctggaagaag accagcccca ggtgctgata tcggatatcc gcatgcccgg cggatcgggc 180ctggacctgc tgcaggccat caaggcgcgg catccgggcc tgccggtcat cgtgatgacg 240gcctactctg acctggacag cgccgtggcc gcgttccagg gcggtgcctt cgaatacctg 300gccaagccct tcgatgtcga caaggcggtt gagctgatcc gccgcgcgct ggaagaaagc 360ctgcgcgagg aagaactgga cgaccgcctc gtcgatgcgc ccgagatcct cggccaggcg 420ccggcgatgc aggacgtgtt ccgcgccatc ggccggctct cgcagtccaa cgtgacggtg 480atgatcaccg gcgagtctgg caccggcaag gagctggtcg cacgcgcgct gcacaagcac 540agcccgcgtg ccaacggtcc ctttatcgcg ctcaataccg cggccatccc caaggacctg 600ctcgaatccg aactgttcgg ccatgagcgc ggcgccttca ccggtgcgca gaccatgcgg 660cgcggccgct tcgagcaggc cgagggcggc acgctgttcc tcgacgaaat cggcgatatg 720ccgttcgacc tgcagacgcg cctgctgcgc gtgctgtccg atggcaactt ctatcgtgtc 780ggcggccaca accccttgcg cgccaatgtg cgcgtgattg ccgccaccca ccagaacctg 840gagctgcgcg tcaaggaagg gctgttccgc gaggacttgt tccaccgcct gaacgtgatc 900cggttgcgcc tgccgccgct gcgcgaacgc ccggaggaca tcacgctgct ggcgcgtcat 960ttcctgcaga agagcgccaa ggaactgggc gtcgagccca agcgcatgtc cgacgaagcg 1020ctggcctatg tcagcacgct gccattcccc ggcaacgtgc gccagctgga gaacctgtgc 1080aactggctga ccgtgatggc gccggcccag accatcgagg tcaaggacct gccgcgcgag 1140atgctggaag ccggcaccag cgagccggtc aatgcgccgc ggcccgagcg cgttgccgaa 1200gcgcgcgcgc ccgagtatga aagcgcgcct gacctggccg actacggcgg ctacgcgacg 1260acggtggcag aggccgacac ggccaccgca gtccggcccg cgtccgtggc gacggtggtc 1320ccggccctgg cctcggccgg ctgggaaagc ctgctcgccg gcgaagcacg ggcaatgctc 1380gaagccggcc agccggaggt catggatgtg ctgacgcgcc gcttcgagaa ggccgtgctg 1440gaggccgcgc tgggtgtcac gcgcgggcgc cgcgtcgagg cggcgacccg gctcgggatc 1500gggcgcaata ccatcacgcg caagctgcag gagcttgggt tcgactga 154840515PRTC. necator 40Met Lys Pro Ile Trp Ile Val Asp Asp Asp Gln Ser Ile Arg Trp Val1 5 10 15Leu Glu Lys Ala Leu Ala Arg Glu Ser Leu Leu Ser Arg Ser Phe Thr 20 25 30Asn Val Arg Asp Ala Leu Ala Ala Leu Glu Glu Asp Gln Pro Gln Val 35 40 45Leu Ile Ser Asp Ile Arg Met Pro Gly Gly Ser Gly Leu Asp Leu Leu 50 55 60Gln Ala Ile Lys Ala Arg His Pro Gly Leu Pro Val Ile Val Met Thr65 70 75 80Ala Tyr Ser Asp Leu Asp Ser Ala Val Ala Ala Phe Gln Gly Gly Ala 85 90 95Phe Glu Tyr Leu Ala Lys Pro Phe Asp Val Asp Lys Ala Val Glu Leu 100 105 110Ile Arg Arg Ala Leu Glu Glu Ser Leu Arg Glu Glu Glu Leu Asp Asp 115 120 125Arg Leu Val Asp Ala Pro Glu Ile Leu Gly Gln Ala Pro Ala Met Gln 130 135 140Asp Val Phe Arg Ala Ile Gly Arg Leu Ser Gln Ser Asn Val Thr Val145 150 155 160Met Ile Thr Gly Glu Ser Gly Thr Gly Lys Glu Leu Val Ala Arg Ala 165 170 175Leu His Lys His Ser Pro Arg Ala Asn Gly Pro Phe Ile Ala Leu Asn 180 185 190Thr Ala Ala Ile Pro Lys Asp Leu Leu Glu Ser Glu Leu Phe Gly His 195 200 205Glu Arg Gly Ala Phe Thr Gly Ala Gln Thr Met Arg Arg Gly Arg Phe 210 215 220Glu Gln Ala Glu Gly Gly Thr Leu Phe Leu Asp Glu Ile Gly Asp Met225 230 235 240Pro Phe Asp Leu Gln Thr Arg Leu Leu Arg Val Leu Ser Asp Gly Asn 245 250 255Phe Tyr Arg Val Gly Gly His Asn Pro Leu Arg Ala Asn Val Arg Val 260 265 270Ile Ala Ala Thr His Gln Asn Leu Glu Leu Arg Val Lys Glu Gly Leu 275 280 285Phe Arg Glu Asp Leu Phe His Arg Leu Asn Val Ile Arg Leu Arg Leu 290 295 300Pro Pro Leu Arg Glu Arg Pro Glu Asp Ile Thr Leu Leu Ala Arg His305 310 315 320Phe Leu Gln Lys Ser Ala Lys Glu Leu Gly Val Glu Pro Lys Arg Met 325 330 335Ser Asp Glu Ala Leu Ala Tyr Val Ser Thr Leu Pro Phe Pro Gly Asn 340 345 350Val Arg Gln Leu Glu Asn Leu Cys Asn Trp Leu Thr Val Met Ala Pro 355 360 365Ala Gln Thr Ile Glu Val Lys Asp Leu Pro Arg Glu Met Leu Glu Ala 370 375 380Gly Thr Ser Glu Pro Val Asn Ala Pro Arg Pro Glu Arg Val Ala Glu385 390 395 400Ala Arg Ala Pro Glu Tyr Glu Ser Ala Pro Asp Leu Ala Asp Tyr Gly 405 410 415Gly Tyr Ala Thr Thr Val Ala Glu Ala Asp Thr Ala Thr Ala Val Arg 420 425 430Pro Ala Ser Val Ala Thr Val Val Pro Ala Leu Ala Ser Ala Gly Trp 435 440 445Glu Ser Leu Leu Ala Gly Glu Ala Arg Ala Met Leu Glu Ala Gly Gln 450 455 460Pro Glu Val Met Asp Val Leu Thr Arg Arg Phe Glu Lys Ala Val Leu465 470 475 480Glu Ala Ala Leu Gly Val Thr Arg Gly Arg Arg Val Glu Ala Ala Thr 485 490 495Arg Leu Gly Ile Gly Arg Asn Thr Ile Thr Arg Lys Leu Gln Glu Leu 500 505 510Gly Phe Asp 515411179DNAC. necator 41atgcgtcgcc tgattcgcgg agtgtcgcgc aaggccagcg gtgccgacgg cgcccgtggc 60gaatcgccgg cggctgatgc ggcgccggcc gaaggcagcg cccaggtgct gaccatgggc 120gccgtggcgt tccatgccgg cctggacgta gtggccaacc cggtgctgct ggtgcagcag 180ccggggctgc gcgtcgtctt tgcgaacccg gccgctgaag ccaccttcgg cgtgtcgcgc 240aagggcatgg tggagctgac gctgcctgac ctgttcgggc gttccgatga gctgcacagc 300atgatcgaca ccgtggtcac gcggcagttc gacgtgcgcc ggcaggacct gatcctgcac 360ccgccgctgc aggagccggc ccatgtgcac gtggtgatct ccgcgctgga agcggtcggc 420gacaccgtgg tggtggaaat cctgcccaat gaacagaagg tgcgcagcga acgcgaagaa 480cgcatcctgg acctgacctc ggccaacaag gaactgatcc gcaacctggc ccacgaaatc 540aagaacccgc tgggcggcat tcgcggcgcg gcgcagctgc tggagttcga gctgccggag 600cgctcgctgc gcgaatacac gcaggtcatc atcaaggaat cggaccggct gcagacgctg 660gtggaccggc tgctggagcc gcaccggcat ccgcatatcg tgtccagcct gaatatccac 720gaagtgctgg agcgcgtgcg ctcggtggtg ctggcggagt tccccaacgg gctggagatc 780gtgcgcgact acgacgccag cctgcccgag ctgcagggcg acatggagca actgatccag 840gccgtgctca acatcgtgca caacgccgcg caggcgctgg ccgaccgcat ggcgcgcggc 900gatgcacaga tcgtgctgcg cacgcgcgtg gcgcgccagg tcacgattgc caagcgcttg 960ttcaagctgg cattggactt gcatgtcatc gacaacggcc cgggaatttc cgaagacatc 1020cgcgaacgca tcttctatcc gctggtatcg ggcagggatg gcggcagcgg actcggtctc 1080acactcgctc aaaccttcgt gcagcagcac gagggcttga tcgaatgcga gagcaggccg 1140ggctgtaccg acttccgcat cctgctgccg ctgcactag 117942392PRTC. necator 42Met Arg Arg Leu Ile Arg Gly Val Ser Arg Lys Ala Ser Gly Ala Asp1 5 10 15Gly Ala Arg Gly Glu Ser Pro Ala Ala Asp Ala Ala Pro Ala Glu Gly 20 25 30Ser Ala Gln Val Leu Thr Met Gly Ala Val Ala Phe His Ala Gly Leu 35 40 45Asp Val Val Ala Asn Pro Val Leu Leu Val Gln Gln Pro Gly Leu Arg 50 55 60Val Val Phe Ala Asn Pro Ala Ala Glu Ala Thr Phe Gly Val Ser Arg65 70 75 80Lys Gly Met Val Glu Leu Thr Leu Pro Asp Leu Phe Gly Arg Ser Asp 85 90 95Glu Leu His Ser Met Ile Asp Thr Val Val Thr Arg Gln Phe Asp Val 100 105 110Arg Arg Gln Asp Leu Ile Leu His Pro Pro Leu Gln Glu Pro Ala His 115 120 125Val His Val Val Ile Ser Ala Leu Glu Ala Val Gly Asp Thr Val Val 130 135 140Val Glu Ile Leu Pro Asn Glu Gln Lys Val Arg Ser Glu Arg Glu Glu145 150 155 160Arg Ile Leu Asp Leu Thr Ser Ala Asn Lys Glu Leu Ile Arg Asn Leu 165 170 175Ala His Glu Ile Lys Asn Pro Leu Gly Gly Ile Arg Gly Ala Ala Gln 180 185 190Leu Leu Glu Phe Glu Leu Pro Glu Arg Ser Leu Arg Glu Tyr Thr Gln 195 200 205Val Ile Ile Lys Glu Ser Asp Arg Leu Gln Thr Leu Val Asp Arg Leu 210 215 220Leu Glu Pro His Arg His Pro His Ile Val Ser Ser Leu Asn Ile His225 230 235 240Glu Val Leu Glu Arg Val Arg Ser Val Val Leu Ala Glu Phe Pro Asn 245 250 255Gly Leu Glu Ile Val Arg Asp Tyr Asp Ala Ser Leu Pro Glu Leu Gln 260 265 270Gly Asp Met Glu Gln Leu Ile Gln Ala Val Leu Asn Ile Val His Asn 275 280 285Ala Ala Gln Ala Leu Ala Asp Arg Met Ala Arg Gly Asp Ala Gln Ile 290 295 300Val Leu Arg Thr Arg Val Ala Arg Gln Val Thr Ile Ala Lys Arg Leu305 310 315 320Phe Lys Leu Ala Leu Asp Leu His Val Ile Asp Asn Gly Pro Gly Ile 325 330 335Ser Glu Asp Ile Arg Glu Arg Ile Phe Tyr Pro Leu Val Ser Gly Arg 340 345 350Asp Gly Gly Ser Gly Leu Gly Leu Thr Leu Ala Gln Thr Phe Val Gln 355 360 365Gln His Glu Gly Leu Ile Glu Cys Glu Ser Arg Pro Gly Cys Thr Asp 370 375 380Phe Arg Ile Leu Leu Pro Leu His385 390431416DNAC. necator 43atggcccaca gcgttgcaga cgtgatgaag ctggtgaagg aaaacgacgt caagttcgtc 60gatttccgtt tcaccgatac caaaggcaag gagcaacacg tgtccgtgcc cgtgtcgcac 120ttcgatgaag acaagttcga aagcggccac gccttcgacg gttcgtcgat cgccggctgg 180aagggtatcg aagcttcaga catgctgctg atgccggatt cgaacaccgc ccacatcgac 240ccgttctacg aagagccgac gctggtgctg tcctgcgacg tggtcgagcc gtcggacggc 300aagggctatg accgcgaccc gcgttccatc gccaagcgcg ccgaagccta cctgaagagc 360accggcctgg gcgacaccgc tttctttggt ccggagcccg agttcttcat cttcgacggc 420gtgacctgga acgtcgacat gcaaggctgc ttcgtgaaga tccattccga agaagccccg 480tggtcgtcgg ccaaggaatt cgagcacggc aacagcggcc accgtccggg caagaagggc 540ggctacttcc cggtcgcccc gatcgacacc ttccaggaca tgcgttcgga aatgtgcctg 600atcctggaat cgctgggcat ccccgttgaa gtccaccacc acgaagtggc tggccagggc 660cagaacgaaa tcggcacccg cttcagcacg ctggtgcagc gcgccgactg gacccagatg 720cagaagtacg tgatccagaa cgtcgcccac acctacggca agaccgccac cttcatgccg 780aagccgatcg ttggcgacaa cggttcgggc atgcacgtgc accagtccgt gtggaaggac 840ggccagaacc tgttcgcggg caacggctac gccggcctgt cggaattcgc gctgtactac 900atcggcggca tcatcaagca cgcccgtgcc ctgaatgcca tcaccaaccc gggcacgaac 960tcgtacaagc gcctggtgcc gggcttcgaa gctccggtga agctggccta ctcggcccgc 1020aaccgctcgg cttcgatccg catcccgtat gtggccaacc cgaagggccg ccgcatcgag 1080acccgcttcc cggatccgct gatgaacccg tacctgggct tctcggcgct gctgatggcc 1140ggcctggatg gcgtgatgaa caagatccac ccgggcgaag ctgccgacaa gaacctgtac 1200gacctgccgc cggaagagga tgcaaagatc ccgaccgtgt gctcgagcct ggaccaggcg 1260ctggagtacc tggacaacga ccgcgagttc ctgacccgcg gcggcgtgtt ctcgaactcg 1320atgatcgatg cctacatcga actgaagatg gaagaagtca cgcgtttccg catgaccacg 1380cacccggtcg agttcgaaat gtactactcg ctgtaa 141644471PRTC. necator 44Met Ala His Ser Val Ala Asp Val Met Lys Leu Val Lys Glu Asn Asp1 5 10 15Val Lys Phe Val Asp Phe Arg Phe Thr Asp Thr Lys Gly Lys Glu Gln 20 25 30His Val Ser Val Pro Val Ser His Phe Asp Glu Asp Lys Phe Glu Ser 35 40 45Gly His Ala Phe Asp Gly Ser Ser Ile Ala Gly Trp Lys Gly Ile Glu 50 55 60Ala Ser Asp Met Leu Leu Met Pro Asp Ser Asn Thr Ala His Ile Asp65 70 75 80Pro Phe Tyr Glu Glu Pro Thr Leu Val Leu Ser Cys Asp Val Val Glu 85 90 95Pro Ser Asp Gly Lys Gly Tyr Asp Arg Asp Pro Arg Ser Ile Ala Lys 100 105 110Arg Ala Glu Ala Tyr Leu Lys Ser Thr Gly Leu Gly Asp Thr Ala Phe 115 120 125Phe Gly Pro Glu Pro Glu Phe Phe Ile Phe Asp Gly Val Thr Trp Asn 130 135 140Val Asp Met Gln Gly Cys Phe Val Lys Ile His Ser Glu Glu Ala Pro145 150 155 160Trp Ser Ser Ala Lys Glu Phe Glu His Gly Asn Ser Gly His Arg Pro 165 170 175Gly Lys Lys Gly Gly Tyr Phe Pro Val Ala Pro Ile Asp Thr Phe Gln 180 185 190Asp Met Arg Ser Glu Met Cys Leu Ile Leu Glu Ser Leu Gly Ile Pro 195 200 205Val Glu Val His His His Glu Val Ala Gly Gln Gly Gln Asn Glu Ile 210 215 220Gly Thr Arg Phe Ser Thr Leu Val Gln Arg Ala Asp Trp Thr Gln Met225 230 235 240Gln Lys Tyr Val Ile Gln Asn Val Ala His Thr Tyr Gly Lys Thr Ala 245 250 255Thr Phe Met Pro Lys Pro Ile Val Gly Asp Asn Gly Ser Gly Met His 260 265 270Val His Gln Ser Val Trp Lys Asp Gly Gln Asn Leu Phe Ala Gly Asn 275 280 285Gly Tyr Ala Gly Leu Ser Glu Phe Ala Leu Tyr Tyr Ile Gly Gly Ile 290 295 300Ile Lys His Ala Arg Ala Leu Asn Ala Ile Thr Asn Pro Gly Thr Asn305 310 315 320Ser Tyr Lys Arg Leu Val Pro Gly Phe Glu Ala Pro Val Lys Leu Ala 325 330 335Tyr Ser Ala Arg Asn Arg Ser Ala Ser Ile Arg Ile Pro Tyr Val Ala 340 345 350Asn Pro Lys Gly Arg Arg Ile Glu Thr Arg Phe Pro Asp Pro Leu Met 355 360 365Asn Pro Tyr Leu Gly Phe Ser Ala Leu Leu Met Ala Gly Leu Asp Gly 370 375 380Val Met Asn Lys Ile His Pro Gly Glu Ala Ala Asp Lys Asn Leu Tyr385 390 395 400Asp Leu Pro Pro Glu Glu Asp Ala Lys Ile Pro Thr Val Cys Ser Ser 405 410 415Leu Asp Gln Ala Leu Glu Tyr Leu Asp Asn Asp Arg Glu Phe Leu Thr 420 425 430Arg Gly Gly Val Phe Ser Asn Ser Met Ile Asp Ala Tyr Ile Glu Leu 435 440 445Lys Met Glu Glu Val Thr Arg Phe Arg Met Thr Thr His Pro Val Glu 450 455 460Phe Glu Met Tyr Tyr Ser Leu465 47045771DNAC. necator 45atgaccctga atccccgcca gacggccctg ctcgaagaag tccgcaccca gggctttgcc 60tccatcgacg aacttgcgcg caaattcggc gtcacgctcc agacggtgcg ccgcgatgtc 120aacctgctgg ccgggaacgg catgctggcg cgcttccatg gcggggtgcg ggtggagggc 180tccaccaccg agaacatcgc ctaccggcag cggcaggtgc tcaacgccga gggcaaggcg 240cgcatcgcgc gcgcggtggc cgctgcggtg cccgagggct gctcgctgat cctgaacatc 300ggcaccacgg tggaagagat agcgcgcgaa ctgatgcacc accgcgggct gcgcgtgatc 360accaacaacc tgaatgtggc caatatcctt gccgacaacc ccgattgcga ggtcatcgtc 420gccggcggcg tgctgcgctc acgcgaccgc ggcatcgtgg gcgaggccac ggtggagttc 480atccgccagt tcaaggtgga tatcgggctg atcggcatct cgggcatcga gaccgacggc 540acgctgcgcg actacgattt ccgcgaggtc aaggtggcgc ggaccatcat cgagcattca 600cgcgaggtgt ggctggcggc ggacgccagc aagttcaatc gccaggcgat ggtggagctg 660gcgcatgtgt cgcaggtcga ccggctcttt accgacgagc cgctggcagc gccgttcgac 720cagatcgtgg ccgacagtgg ggtgaagtgt gtggtggcgg agcgggagtg a 77146256PRTC. necator 46Met Thr Leu Asn Pro Arg Gln Thr Ala Leu Leu Glu Glu Val Arg Thr1 5 10 15Gln Gly Phe Ala Ser Ile Asp Glu Leu Ala Arg Lys Phe Gly Val Thr 20 25 30Leu Gln Thr Val Arg Arg Asp Val Asn Leu Leu Ala Gly Asn Gly Met 35 40 45Leu Ala Arg Phe His Gly Gly Val Arg Val Glu Gly Ser Thr Thr Glu 50 55 60Asn Ile Ala Tyr Arg Gln Arg Gln Val Leu Asn Ala Glu Gly Lys Ala65 70 75 80Arg Ile Ala Arg Ala Val Ala Ala Ala Val Pro Glu Gly Cys Ser Leu 85 90 95Ile Leu Asn Ile Gly Thr Thr Val Glu Glu Ile Ala Arg Glu Leu Met 100 105 110His His Arg Gly Leu Arg Val Ile Thr Asn Asn Leu Asn Val Ala Asn 115 120 125Ile Leu Ala Asp Asn Pro Asp Cys Glu Val Ile Val Ala Gly Gly Val 130 135 140Leu Arg Ser Arg Asp Arg Gly Ile Val Gly Glu Ala Thr Val Glu Phe145 150 155 160Ile Arg Gln Phe Lys Val Asp Ile Gly Leu Ile Gly Ile Ser Gly Ile 165 170 175Glu Thr Asp Gly Thr Leu Arg

Asp Tyr Asp Phe Arg Glu Val Lys Val 180 185 190Ala Arg Thr Ile Ile Glu His Ser Arg Glu Val Trp Leu Ala Ala Asp 195 200 205Ala Ser Lys Phe Asn Arg Gln Ala Met Val Glu Leu Ala His Val Ser 210 215 220Gln Val Asp Arg Leu Phe Thr Asp Glu Pro Leu Ala Ala Pro Phe Asp225 230 235 240Gln Ile Val Ala Asp Ser Gly Val Lys Cys Val Val Ala Glu Arg Glu 245 250 255471464DNAC. necator 47atgggtaagg cgactggctt tctcgaattt ccgcgccaga acgaaggcta cgaaccggta 60gtcaagcgcg tgaagcacta caaggaattc gtgttcgcgc tgtccgacag cgaagcgaag 120atccagggtg cgcgctgcat ggactgcggc atcccgttct gcaacaacgg ctgcccggtc 180aacaacatca tccccgactt caacgacctg gtgtaccgcc aggactggaa gtcggcgatc 240gaggtgctgc accagaccaa caacttcccc gagttcaccg gccgcatctg ccccgcaccg 300tgcgaggccg cctgcacgct gggcatcaat gaactgccgg tgggcatcaa gtcgatcgag 360cacgccatca tcgacaaggc ctgggaagag ggctgggtca ggccgcagct gccgcgccac 420aagaccggca agaccgtggc cgtggtcggc tccggtcccg ccggcatggc tgccgcgcag 480cagctggcac gcgccggcca tgacgtgacc gtgttcgaga agaacgaccg catcggcggc 540ctgctgcgct acggcatccc cgacttcaag atggagaaga cgctgatcga ccgccgcatc 600gagcagatgc aggccgaagg cgtgaccttc cgtccgggcg tgatggtgac cgacggcgaa 660ctgccggccg gcatcaagaa ctacgcccgt gaaaccatct cggcccaggc cctgatggac 720cagttcgacg ccgtggtgct ggcgggcggc tcggaagtgc cgcgcgacct gccggtgccg 780ggccgcgacc tggccggcat ccacttcgcg ctggaattcc tgatcccgca gaacaaggaa 840gtggcaggcg acggcgaaaa cgagatccgc gccgaaggca agaacgtgat tgtgatcggt 900ggcggcgata ccggctccga ctgcgtgggt acgtccaacc gccatggcgc cacctcggtg 960acgcagtttg aactgctgcc gcagccgccg gaagaagagg acaagccgct ggtgtggccg 1020tactggccga tcaagctgcg cacctcgtcg tcgcacgatg aaggctgcga gcgagactgg 1080tcggtcgcca ccaaggaatt catcggcgag aacggcaagg tcaccgcact gaaggcctgc 1140cgtgtcgaat ggaaggatgg ccgcatgcag gaagtcgaag gcagcgagtt catcctgccg 1200gccgacctgg tgctgctggc gatgggcttt accaacccgg tgggttcgat gctggaagcg 1260tttggcgtgg ataccgatgc gcgcaagaac gccaaggcct cgaccgaggg cgagcgtgcc 1320taccacacca acgtgcccaa ggtgttcgcc gctggcgacg tgcgccgtgg ccagtcgctg 1380gtggtgtggg cgatccgcga aggccgccag gccgcgcgtt cggtcgatgc cttcctgatg 1440ggtcacaccg aactgccgcg ctga 146448487PRTC. necator 48Met Gly Lys Ala Thr Gly Phe Leu Glu Phe Pro Arg Gln Asn Glu Gly1 5 10 15Tyr Glu Pro Val Val Lys Arg Val Lys His Tyr Lys Glu Phe Val Phe 20 25 30Ala Leu Ser Asp Ser Glu Ala Lys Ile Gln Gly Ala Arg Cys Met Asp 35 40 45Cys Gly Ile Pro Phe Cys Asn Asn Gly Cys Pro Val Asn Asn Ile Ile 50 55 60Pro Asp Phe Asn Asp Leu Val Tyr Arg Gln Asp Trp Lys Ser Ala Ile65 70 75 80Glu Val Leu His Gln Thr Asn Asn Phe Pro Glu Phe Thr Gly Arg Ile 85 90 95Cys Pro Ala Pro Cys Glu Ala Ala Cys Thr Leu Gly Ile Asn Glu Leu 100 105 110Pro Val Gly Ile Lys Ser Ile Glu His Ala Ile Ile Asp Lys Ala Trp 115 120 125Glu Glu Gly Trp Val Arg Pro Gln Leu Pro Arg His Lys Thr Gly Lys 130 135 140Thr Val Ala Val Val Gly Ser Gly Pro Ala Gly Met Ala Ala Ala Gln145 150 155 160Gln Leu Ala Arg Ala Gly His Asp Val Thr Val Phe Glu Lys Asn Asp 165 170 175Arg Ile Gly Gly Leu Leu Arg Tyr Gly Ile Pro Asp Phe Lys Met Glu 180 185 190Lys Thr Leu Ile Asp Arg Arg Ile Glu Gln Met Gln Ala Glu Gly Val 195 200 205Thr Phe Arg Pro Gly Val Met Val Thr Asp Gly Glu Leu Pro Ala Gly 210 215 220Ile Lys Asn Tyr Ala Arg Glu Thr Ile Ser Ala Gln Ala Leu Met Asp225 230 235 240Gln Phe Asp Ala Val Val Leu Ala Gly Gly Ser Glu Val Pro Arg Asp 245 250 255Leu Pro Val Pro Gly Arg Asp Leu Ala Gly Ile His Phe Ala Leu Glu 260 265 270Phe Leu Ile Pro Gln Asn Lys Glu Val Ala Gly Asp Gly Glu Asn Glu 275 280 285Ile Arg Ala Glu Gly Lys Asn Val Ile Val Ile Gly Gly Gly Asp Thr 290 295 300Gly Ser Asp Cys Val Gly Thr Ser Asn Arg His Gly Ala Thr Ser Val305 310 315 320Thr Gln Phe Glu Leu Leu Pro Gln Pro Pro Glu Glu Glu Asp Lys Pro 325 330 335Leu Val Trp Pro Tyr Trp Pro Ile Lys Leu Arg Thr Ser Ser Ser His 340 345 350Asp Glu Gly Cys Glu Arg Asp Trp Ser Val Ala Thr Lys Glu Phe Ile 355 360 365Gly Glu Asn Gly Lys Val Thr Ala Leu Lys Ala Cys Arg Val Glu Trp 370 375 380Lys Asp Gly Arg Met Gln Glu Val Glu Gly Ser Glu Phe Ile Leu Pro385 390 395 400Ala Asp Leu Val Leu Leu Ala Met Gly Phe Thr Asn Pro Val Gly Ser 405 410 415Met Leu Glu Ala Phe Gly Val Asp Thr Asp Ala Arg Lys Asn Ala Lys 420 425 430Ala Ser Thr Glu Gly Glu Arg Ala Tyr His Thr Asn Val Pro Lys Val 435 440 445Phe Ala Ala Gly Asp Val Arg Arg Gly Gln Ser Leu Val Val Trp Ala 450 455 460Ile Arg Glu Gly Arg Gln Ala Ala Arg Ser Val Asp Ala Phe Leu Met465 470 475 480Gly His Thr Glu Leu Pro Arg 485494806DNAC. necator 49gtggaccaaa cgaaaaatct ttcggcccaa gctcaggcac aagcgcagac cagcgagtct 60tcccacgcca ttgacctgcg cccgcaagcg cagggcatgt acgatcccag caacgagcat 120gacgcctgcg gcgtcggcat ggtcgcgcat atcaagggca agaagtccca cgagatcatc 180tcgcagggcc tgaagatcct ggagaacctg gaccaccggg gtgcggtcgg cgccgatgcg 240ctgatgggcg acggtgccgg catcctgatc cagatcccgg accagttcta ccgcgaggaa 300atggccgcgc agggcgtgag cctgccgccc gccggcgaat acggcgtggg catgatcttc 360ctgccgaagg aacacgcctc gcgcctggcc tgcgagcagg aactggagcg cacggtccgc 420ctggaaggcc aggtcgtgct gggctggcgc gacgtgccgg tcgacgccaa gatgccgatg 480tcgccgacgg tgcgcaccac cgagccggtg atccgccaga tcttcatcgg gcgcggccgc 540gacatcatga ccacggacgc gctggaacgt aagctctacg tcatccgcaa gaccgccagc 600catgccatcc aggcgctcaa gctcaagcac ggcaaggaat acttcgtgcc gtcaatgtcg 660gcccgtaccg tggtgtacaa gggcctgctg ctggccaacc aggttggcga gtactacctg 720gacctgctgg accagcgcgc cgtctcggcc ctggccctgg tgcaccagcg cttctcgacc 780aacaccttcc cggcctggga actggcccac ccgtaccgca tggtcgccca caacggcgaa 840atcaacacgg tcaagggcaa tgtcaactgg atcaacgcgc gcaccggcgc gatctcgtcg 900ccggtgctcg gcgacgacct gcccaagctg tggccgctga tctacccggg ccagtccgac 960accgcatcgt tcgacaactg cctcgaactg ctgacgatgg ccggctaccc gctcgtccac 1020gcgatgatga tgatgatccc ggaagcctgg gaacagcaca cgctgatgga cgacaaccgc 1080cgcgccttct acgagtacca cgccgccatg atggagccgt gggacggccc ggccgcgatc 1140tgcttcaccg atggccgcca gatcggcgcc acgctggacc gcaacggcct gcgcccggca 1200cgtttctacg tgaccgagga cgacatcgtg gtgctggctt cggaagccgg cgtgctgccg 1260ttccccgagt cgcgcatcgt tgagaagtgg cgcctgcagc cgggcaagat gttcctgatc 1320gacatggaac agggccgcat catcgacgac aaggaactca aggacaacct ggccaacgcc 1380aagccgtaca agagctggat cgacgccgtg cgcatcaagc tcgacgagct cgacgccaag 1440cctgaagacg ttgccgccga gaagaagccc gtggccaagc tgctggaccg ccagcaggcc 1500tttggctaca cccaggaaga cgtcaagttc ctgatggcgc cgatggcgct ggccggcgag 1560gaagccaccg gctcgatggg caacgattcg ccgctggcca tcctgtcgtc caagaacaag 1620acgctgtacc actacttcaa gcagctgttc gcccaggtca ccaacccgcc gatcgacccg 1680atccgcgaga acatggtgat gtcgctggtg tcgttcatcg gcccgaagcc gaacctgctc 1740gagctgaaca acatcaaccc gccgatgcgc ctcgaagtgt cccagccggt gctggacttc 1800aaggacatcg ccaagatccg caatatcgag cactacaccg gcggcaagtt ccgttcgtac 1860gagctgaaca tctgctaccc gaaggcctgg ggcaaggagg gcatcgaagc gcgcctggcc 1920tcgctgtgcg ccgaagccgt ggatgcggtg cgttcgggct tcaacatcct gatcgtgtcg 1980gaccgccgcg tggatgccga gcatgttgcg attcccgcgc tgctggccac gtccgccatc 2040caccaccacc tggtggagaa gggcctgcgc acgtccaccg gcctggtggt cgagaccggc 2100accgcccgtg aagtgcacca cttcgcgctg ctggccggct atggcgccga agccgtgcat 2160ccgtacctgg cgatggaaac gctggccgaa atggcccagg gcctgtccgg cgacctgtcg 2220cccgagaagg cggtcaagaa cttcgtcaag gcgatcggca agggcctgtt caaagtgatg 2280tccaagatgg gcatctccac ctacatgtcg tacaccggcg cgcagatctt cgaagccatc 2340ggcctgtcgc gcgaactggt gcagaagtac ttccatggca ccccgtcgaa tgtcgagggc 2400atcggcatct tcgaagtggc cgaggaagcc ctgcgcctgc accgcgacgc ctttggcgac 2460aacccggtgc tggaaagcat gctggacgcc ggcggcgaat acgccttccg catccgcggc 2520gaagagcata tgtggacccc ggactcggtc gccaagctgc agcactcggt gcgcgccgac 2580gacggcaagg gcgcctacca gacgtacaag gaatacgcca acatcatcaa cgaccagagc 2640aagcgccaca tgacgctgcg tggcctgttc gagttcaagg tcgatccggc caaggcgatt 2700ccgctggaag aggtggagtc ggccaaggag atcgtcaagc gcttcgccac cggtgcgatg 2760tcgctcggct cgatctcgac cgaagcccac accacgctgg cgctggcgat gaaccgcatc 2820ggcggcaagt ccaacaccgg cgaaggcggc gaggacgaga agcgctaccg caacgagctg 2880cgcggcattc ccatcaagca gggcgatacc ctcaagggcc tgctgggcga caacgtgatc 2940gaacgcgacc tggaactgca ggaaggcgat tcgctgcgct cgaagatcaa gcaggtggcg 3000tcgggccgtt tcggcgtgac cgccgaatac ctggcttcgg ccgaccagat ccagatcaag 3060atggcgcagg gtgccaagcc cggcgaaggc ggccagctgc ccggccacaa ggtctcggac 3120tacatcggca agctgcgtta ctcggtgccg ggcgtgggcc tgatctcgcc gcccccgcac 3180catgacatct attcgatcga ggatctggca cagctgatcc acgacctgaa gaacgtcaac 3240ccggtgtcgg acatctcggt caagctggtg tccgaagtcg gcgtcggcac ggtggccgcg 3300ggcgtggcca aggccaaggc cgaccacgtc gtgatcgccg gccatgatgg cggcaccggc 3360gcttcgccgt ggtcgtcgat caagcatgcc ggcacgccgt gggagctggg cctggccgaa 3420acgcagcaga cgctgctgct caacggcctg cgcaaccgca tccgcgtgca ggccgacggc 3480cagatgaaga ccggccgcga cgtcgtcatc ggcgcgctgc tgggcgccga tgaattcggc 3540ttcgccaccg cgccgctggt tgcggaaggc tgcatcatga tgcgcaagtg ccacctgaac 3600acctgcccgg tgggcgtggc cacgcaggat ccgcagctgc gcaggaagtt ccagggcaag 3660cctgagcacg tggtcaactt cttcttcttc gttgcggaag aagcccgcga aatcatggcc 3720cagctgggca tccgcaagtt cgacgagctg atcggccgcg ccgacctgct cgacaccaag 3780cccggcatcg agcactggaa ggcgcgcggc ctggacttcg gccgcatctt ccaccaggtc 3840tcgctgggcg cggacgtgcc gctgtaccac accgacgtgc aggaccacgg cctgtcggcc 3900gaggccggca aggcgctgga ccacgtgctg atcgccaagg cccgaccggc gatcgagaag 3960ggcgagcggg tctcgttcat ccagccggtg aagaacgtca accgtaccgt cggcgcgatg 4020ctgtcgggcg tggtggcgcg ccagcatggc cacgaaggcc tgcctgacga taccatccac 4080atccagctgc aaggcaccgc cggccagtcg ttcggcgcgt tcctggcgca cggcatcacg 4140ctggacctgg tgggcgacgg caacgactat gtcggcaagg gcctgtcggg cggccgcgtg 4200atcgtgcgcg ctccgcatga gttccgcggc gacccgaccc gcaacatcat cgtcggcaac 4260accgtgctgt acggtgctat cgccggggaa gcgttcttca acggcgtggc cggcgagcgc 4320ttcgcggtgc gcaactcggg tgcggtggca gtggtggaag gcaccggcga ccacggttgc 4380gagtacatga ccggcggcac ggtggtggtg ctgggcggca ccggacgcaa cttcgcggcc 4440ggcatgtcgg gcggcgtggc ctacgtctac gacgaggacg gcctgttcga caagcgctgc 4500aacacctcga tggtggcgct ggaagcagtg ctggcttcgg ccgaccagga gaagggccag 4560cccgaggctt cgtggcacaa ggtcgacggc aagcgccagc tggatgaggt catcctgcgc 4620aacctgatcg agcagcattt ccgctacacc ggttccgagc gcgccaaggc gctgctggcc 4680gactggacca cggcacgccg caagttcgtc aaggtcttcc cgaccgagta caagcgcgcg 4740ctgggcgaga tgtacgccaa ggaacaggcc gcccgcgaca gcgaccgcga agccatcgcg 4800gcctga 4806501601PRTC. necator 50Met Asp Gln Thr Lys Asn Leu Ser Ala Gln Ala Gln Ala Gln Ala Gln1 5 10 15Thr Ser Glu Ser Ser His Ala Ile Asp Leu Arg Pro Gln Ala Gln Gly 20 25 30Met Tyr Asp Pro Ser Asn Glu His Asp Ala Cys Gly Val Gly Met Val 35 40 45Ala His Ile Lys Gly Lys Lys Ser His Glu Ile Ile Ser Gln Gly Leu 50 55 60Lys Ile Leu Glu Asn Leu Asp His Arg Gly Ala Val Gly Ala Asp Ala65 70 75 80Leu Met Gly Asp Gly Ala Gly Ile Leu Ile Gln Ile Pro Asp Gln Phe 85 90 95Tyr Arg Glu Glu Met Ala Ala Gln Gly Val Ser Leu Pro Pro Ala Gly 100 105 110Glu Tyr Gly Val Gly Met Ile Phe Leu Pro Lys Glu His Ala Ser Arg 115 120 125Leu Ala Cys Glu Gln Glu Leu Glu Arg Thr Val Arg Leu Glu Gly Gln 130 135 140Val Val Leu Gly Trp Arg Asp Val Pro Val Asp Ala Lys Met Pro Met145 150 155 160Ser Pro Thr Val Arg Thr Thr Glu Pro Val Ile Arg Gln Ile Phe Ile 165 170 175Gly Arg Gly Arg Asp Ile Met Thr Thr Asp Ala Leu Glu Arg Lys Leu 180 185 190Tyr Val Ile Arg Lys Thr Ala Ser His Ala Ile Gln Ala Leu Lys Leu 195 200 205Lys His Gly Lys Glu Tyr Phe Val Pro Ser Met Ser Ala Arg Thr Val 210 215 220Val Tyr Lys Gly Leu Leu Leu Ala Asn Gln Val Gly Glu Tyr Tyr Leu225 230 235 240Asp Leu Leu Asp Gln Arg Ala Val Ser Ala Leu Ala Leu Val His Gln 245 250 255Arg Phe Ser Thr Asn Thr Phe Pro Ala Trp Glu Leu Ala His Pro Tyr 260 265 270Arg Met Val Ala His Asn Gly Glu Ile Asn Thr Val Lys Gly Asn Val 275 280 285Asn Trp Ile Asn Ala Arg Thr Gly Ala Ile Ser Ser Pro Val Leu Gly 290 295 300Asp Asp Leu Pro Lys Leu Trp Pro Leu Ile Tyr Pro Gly Gln Ser Asp305 310 315 320Thr Ala Ser Phe Asp Asn Cys Leu Glu Leu Leu Thr Met Ala Gly Tyr 325 330 335Pro Leu Val His Ala Met Met Met Met Ile Pro Glu Ala Trp Glu Gln 340 345 350His Thr Leu Met Asp Asp Asn Arg Arg Ala Phe Tyr Glu Tyr His Ala 355 360 365Ala Met Met Glu Pro Trp Asp Gly Pro Ala Ala Ile Cys Phe Thr Asp 370 375 380Gly Arg Gln Ile Gly Ala Thr Leu Asp Arg Asn Gly Leu Arg Pro Ala385 390 395 400Arg Phe Tyr Val Thr Glu Asp Asp Ile Val Val Leu Ala Ser Glu Ala 405 410 415Gly Val Leu Pro Phe Pro Glu Ser Arg Ile Val Glu Lys Trp Arg Leu 420 425 430Gln Pro Gly Lys Met Phe Leu Ile Asp Met Glu Gln Gly Arg Ile Ile 435 440 445Asp Asp Lys Glu Leu Lys Asp Asn Leu Ala Asn Ala Lys Pro Tyr Lys 450 455 460Ser Trp Ile Asp Ala Val Arg Ile Lys Leu Asp Glu Leu Asp Ala Lys465 470 475 480Pro Glu Asp Val Ala Ala Glu Lys Lys Pro Val Ala Lys Leu Leu Asp 485 490 495Arg Gln Gln Ala Phe Gly Tyr Thr Gln Glu Asp Val Lys Phe Leu Met 500 505 510Ala Pro Met Ala Leu Ala Gly Glu Glu Ala Thr Gly Ser Met Gly Asn 515 520 525Asp Ser Pro Leu Ala Ile Leu Ser Ser Lys Asn Lys Thr Leu Tyr His 530 535 540Tyr Phe Lys Gln Leu Phe Ala Gln Val Thr Asn Pro Pro Ile Asp Pro545 550 555 560Ile Arg Glu Asn Met Val Met Ser Leu Val Ser Phe Ile Gly Pro Lys 565 570 575Pro Asn Leu Leu Glu Leu Asn Asn Ile Asn Pro Pro Met Arg Leu Glu 580 585 590Val Ser Gln Pro Val Leu Asp Phe Lys Asp Ile Ala Lys Ile Arg Asn 595 600 605Ile Glu His Tyr Thr Gly Gly Lys Phe Arg Ser Tyr Glu Leu Asn Ile 610 615 620Cys Tyr Pro Lys Ala Trp Gly Lys Glu Gly Ile Glu Ala Arg Leu Ala625 630 635 640Ser Leu Cys Ala Glu Ala Val Asp Ala Val Arg Ser Gly Phe Asn Ile 645 650 655Leu Ile Val Ser Asp Arg Arg Val Asp Ala Glu His Val Ala Ile Pro 660 665 670Ala Leu Leu Ala Thr Ser Ala Ile His His His Leu Val Glu Lys Gly 675 680 685Leu Arg Thr Ser Thr Gly Leu Val Val Glu Thr Gly Thr Ala Arg Glu 690 695 700Val His His Phe Ala Leu Leu Ala Gly Tyr Gly Ala Glu Ala Val His705 710 715 720Pro Tyr Leu Ala Met Glu Thr Leu Ala Glu Met Ala Gln Gly Leu Ser 725 730 735Gly Asp Leu Ser Pro Glu Lys Ala Val Lys Asn Phe Val Lys Ala Ile 740 745 750Gly Lys Gly Leu Phe Lys Val Met Ser Lys Met Gly Ile Ser Thr Tyr 755 760 765Met Ser Tyr Thr Gly Ala Gln Ile Phe Glu Ala Ile Gly Leu Ser Arg 770 775 780Glu Leu Val Gln Lys Tyr Phe His Gly Thr Pro Ser Asn Val Glu Gly785 790 795 800Ile Gly Ile Phe Glu Val Ala Glu Glu Ala Leu Arg Leu His Arg Asp 805 810 815Ala Phe Gly Asp Asn Pro Val Leu Glu Ser Met Leu Asp Ala Gly Gly 820 825 830Glu Tyr Ala Phe Arg Ile Arg Gly Glu Glu His Met Trp Thr Pro Asp 835 840 845Ser Val Ala Lys Leu Gln His Ser Val Arg Ala Asp

Asp Gly Lys Gly 850 855 860Ala Tyr Gln Thr Tyr Lys Glu Tyr Ala Asn Ile Ile Asn Asp Gln Ser865 870 875 880Lys Arg His Met Thr Leu Arg Gly Leu Phe Glu Phe Lys Val Asp Pro 885 890 895Ala Lys Ala Ile Pro Leu Glu Glu Val Glu Ser Ala Lys Glu Ile Val 900 905 910Lys Arg Phe Ala Thr Gly Ala Met Ser Leu Gly Ser Ile Ser Thr Glu 915 920 925Ala His Thr Thr Leu Ala Leu Ala Met Asn Arg Ile Gly Gly Lys Ser 930 935 940Asn Thr Gly Glu Gly Gly Glu Asp Glu Lys Arg Tyr Arg Asn Glu Leu945 950 955 960Arg Gly Ile Pro Ile Lys Gln Gly Asp Thr Leu Lys Gly Leu Leu Gly 965 970 975Asp Asn Val Ile Glu Arg Asp Leu Glu Leu Gln Glu Gly Asp Ser Leu 980 985 990Arg Ser Lys Ile Lys Gln Val Ala Ser Gly Arg Phe Gly Val Thr Ala 995 1000 1005Glu Tyr Leu Ala Ser Ala Asp Gln Ile Gln Ile Lys Met Ala Gln 1010 1015 1020Gly Ala Lys Pro Gly Glu Gly Gly Gln Leu Pro Gly His Lys Val 1025 1030 1035Ser Asp Tyr Ile Gly Lys Leu Arg Tyr Ser Val Pro Gly Val Gly 1040 1045 1050Leu Ile Ser Pro Pro Pro His His Asp Ile Tyr Ser Ile Glu Asp 1055 1060 1065Leu Ala Gln Leu Ile His Asp Leu Lys Asn Val Asn Pro Val Ser 1070 1075 1080Asp Ile Ser Val Lys Leu Val Ser Glu Val Gly Val Gly Thr Val 1085 1090 1095Ala Ala Gly Val Ala Lys Ala Lys Ala Asp His Val Val Ile Ala 1100 1105 1110Gly His Asp Gly Gly Thr Gly Ala Ser Pro Trp Ser Ser Ile Lys 1115 1120 1125His Ala Gly Thr Pro Trp Glu Leu Gly Leu Ala Glu Thr Gln Gln 1130 1135 1140Thr Leu Leu Leu Asn Gly Leu Arg Asn Arg Ile Arg Val Gln Ala 1145 1150 1155Asp Gly Gln Met Lys Thr Gly Arg Asp Val Val Ile Gly Ala Leu 1160 1165 1170Leu Gly Ala Asp Glu Phe Gly Phe Ala Thr Ala Pro Leu Val Ala 1175 1180 1185Glu Gly Cys Ile Met Met Arg Lys Cys His Leu Asn Thr Cys Pro 1190 1195 1200Val Gly Val Ala Thr Gln Asp Pro Gln Leu Arg Arg Lys Phe Gln 1205 1210 1215Gly Lys Pro Glu His Val Val Asn Phe Phe Phe Phe Val Ala Glu 1220 1225 1230Glu Ala Arg Glu Ile Met Ala Gln Leu Gly Ile Arg Lys Phe Asp 1235 1240 1245Glu Leu Ile Gly Arg Ala Asp Leu Leu Asp Thr Lys Pro Gly Ile 1250 1255 1260Glu His Trp Lys Ala Arg Gly Leu Asp Phe Gly Arg Ile Phe His 1265 1270 1275Gln Val Ser Leu Gly Ala Asp Val Pro Leu Tyr His Thr Asp Val 1280 1285 1290Gln Asp His Gly Leu Ser Ala Glu Ala Gly Lys Ala Leu Asp His 1295 1300 1305Val Leu Ile Ala Lys Ala Arg Pro Ala Ile Glu Lys Gly Glu Arg 1310 1315 1320Val Ser Phe Ile Gln Pro Val Lys Asn Val Asn Arg Thr Val Gly 1325 1330 1335Ala Met Leu Ser Gly Val Val Ala Arg Gln His Gly His Glu Gly 1340 1345 1350Leu Pro Asp Asp Thr Ile His Ile Gln Leu Gln Gly Thr Ala Gly 1355 1360 1365Gln Ser Phe Gly Ala Phe Leu Ala His Gly Ile Thr Leu Asp Leu 1370 1375 1380Val Gly Asp Gly Asn Asp Tyr Val Gly Lys Gly Leu Ser Gly Gly 1385 1390 1395Arg Val Ile Val Arg Ala Pro His Glu Phe Arg Gly Asp Pro Thr 1400 1405 1410Arg Asn Ile Ile Val Gly Asn Thr Val Leu Tyr Gly Ala Ile Ala 1415 1420 1425Gly Glu Ala Phe Phe Asn Gly Val Ala Gly Glu Arg Phe Ala Val 1430 1435 1440Arg Asn Ser Gly Ala Val Ala Val Val Glu Gly Thr Gly Asp His 1445 1450 1455Gly Cys Glu Tyr Met Thr Gly Gly Thr Val Val Val Leu Gly Gly 1460 1465 1470Thr Gly Arg Asn Phe Ala Ala Gly Met Ser Gly Gly Val Ala Tyr 1475 1480 1485Val Tyr Asp Glu Asp Gly Leu Phe Asp Lys Arg Cys Asn Thr Ser 1490 1495 1500Met Val Ala Leu Glu Ala Val Leu Ala Ser Ala Asp Gln Glu Lys 1505 1510 1515Gly Gln Pro Glu Ala Ser Trp His Lys Val Asp Gly Lys Arg Gln 1520 1525 1530Leu Asp Glu Val Ile Leu Arg Asn Leu Ile Glu Gln His Phe Arg 1535 1540 1545Tyr Thr Gly Ser Glu Arg Ala Lys Ala Leu Leu Ala Asp Trp Thr 1550 1555 1560Thr Ala Arg Arg Lys Phe Val Lys Val Phe Pro Thr Glu Tyr Lys 1565 1570 1575Arg Ala Leu Gly Glu Met Tyr Ala Lys Glu Gln Ala Ala Arg Asp 1580 1585 1590Ser Asp Arg Glu Ala Ile Ala Ala 1595 160051975DNAC. necator 51atgaagaccg gcaaaagctc ggcaggagac accaaggccc ggattctcga cgccacggaa 60aagttgttta ccgaggtcgg ctatgaagcc acctcgctca ggcaggtcac gtcgcgcgcc 120atcgtcaacc tggccgccgt gaactaccac ttccgcagca aagatatcat gatgcacgcc 180gtgctgagcc ggcggctgga tccgctcaac gcgcggcgcc tggcgctgct cgatgcgtgc 240gaagcgcgct ggcccggcaa cagcatccgc tgcgagcacg tgatgggtgc gctgttcgtg 300cccgcgctgc aaatggcacg cgacccgtcg gtgggcgggc cgtcgttcct gcggctgctc 360gggcgcgtgt attcggatac ctcgcccttt atccagcaat acctgctgga gcactacgcc 420ccggtgtacg ggcgcttctt cgacgccttc tcgcgcgcca ttccggcgct gccgcggcat 480gagctcggct ggcgcctgca gtttgcgctc aaggcgctgg ccggcgtgct ggccggcgaa 540gagctcacca acctgctgcc cgcgttcacg cagggccggc agatgagcga tgcgcacgtg 600ctggcccagc tcaccgccat ggtggaagcc gtgctgaacg tggcgcaacc cagtgcggac 660gacttctcgg ccctgcagtc ggtgttcgag cttggcgagc agcagcaggc cagggagcgc 720gccagcaggc tggccgcggc gcggtctgcc gatgaactga gtgccagttg cgccgcgatg 780gcggcaacca tgggcgatgc ggccagccag accgccgccg ccctcggcaa ggcccgcatc 840agcgcacgca ccgtgcgcaa tgagggcacc aaacgctcag ccgtgaccag catgccggtc 900cgcgcgcgcg agcaaaccgt gagcttcccc agcaacccgc tggatgactg gatgcgaatg 960cgcaccagga catag 97552324PRTC. necator 52Met Lys Thr Gly Lys Ser Ser Ala Gly Asp Thr Lys Ala Arg Ile Leu1 5 10 15Asp Ala Thr Glu Lys Leu Phe Thr Glu Val Gly Tyr Glu Ala Thr Ser 20 25 30Leu Arg Gln Val Thr Ser Arg Ala Ile Val Asn Leu Ala Ala Val Asn 35 40 45Tyr His Phe Arg Ser Lys Asp Ile Met Met His Ala Val Leu Ser Arg 50 55 60Arg Leu Asp Pro Leu Asn Ala Arg Arg Leu Ala Leu Leu Asp Ala Cys65 70 75 80Glu Ala Arg Trp Pro Gly Asn Ser Ile Arg Cys Glu His Val Met Gly 85 90 95Ala Leu Phe Val Pro Ala Leu Gln Met Ala Arg Asp Pro Ser Val Gly 100 105 110Gly Pro Ser Phe Leu Arg Leu Leu Gly Arg Val Tyr Ser Asp Thr Ser 115 120 125Pro Phe Ile Gln Gln Tyr Leu Leu Glu His Tyr Ala Pro Val Tyr Gly 130 135 140Arg Phe Phe Asp Ala Phe Ser Arg Ala Ile Pro Ala Leu Pro Arg His145 150 155 160Glu Leu Gly Trp Arg Leu Gln Phe Ala Leu Lys Ala Leu Ala Gly Val 165 170 175Leu Ala Gly Glu Glu Leu Thr Asn Leu Leu Pro Ala Phe Thr Gln Gly 180 185 190Arg Gln Met Ser Asp Ala His Val Leu Ala Gln Leu Thr Ala Met Val 195 200 205Glu Ala Val Leu Asn Val Ala Gln Pro Ser Ala Asp Asp Phe Ser Ala 210 215 220Leu Gln Ser Val Phe Glu Leu Gly Glu Gln Gln Gln Ala Arg Glu Arg225 230 235 240Ala Ser Arg Leu Ala Ala Ala Arg Ser Ala Asp Glu Leu Ser Ala Ser 245 250 255Cys Ala Ala Met Ala Ala Thr Met Gly Asp Ala Ala Ser Gln Thr Ala 260 265 270Ala Ala Leu Gly Lys Ala Arg Ile Ser Ala Arg Thr Val Arg Asn Glu 275 280 285Gly Thr Lys Arg Ser Ala Val Thr Ser Met Pro Val Arg Ala Arg Glu 290 295 300Gln Thr Val Ser Phe Pro Ser Asn Pro Leu Asp Asp Trp Met Arg Met305 310 315 320Arg Thr Arg Thr53672DNAC. necator 53atgcgccgga tcgagaccgc gctgctcgac ggcacctggc cgcccggcac gcggctgccg 60gccgagcggg tgctggccgg gcaatacgag gtggcgcgca acaccgtgcg cgaggcgatc 120cagcgcctgg ccgcgcgcgg cctgctgcag agccggcgcg gtgccggggt ctacgccacc 180gaccagctgc gcgccggcat tgcctcgccc tggggccagc tggtggccga ccaccccgcg 240ctgcgcgacg acatcctgga gttccgccgc gtgctggagg gcgcgaccgc gtattttgcg 300gcgctgcgcg cggatgccgc tgacgtgaag cggatccgcg ccctgatggc cgagctggaa 360cgcgcgcgcg ccgccgacga caagcaggca gaggccgatg ccgatgcgca gctgcacgac 420gccattgccc aggcctcgca caacaccatg ttcctgcacc tgcataccag cgtgatcggc 480atgctgcgcg agcacatcac catcaatggc accggcctgc gcgaacagga cgacggcgcg 540tcagacctgt tgctgctgca gcatcgcacg ctgtgcgatg cgatttgcgc gcgccgcccg 600gaagaggcgc gcaccgccat gcagacccat atcgacttcg tgcgcagccg ggtggagcag 660gacggcgcct ga 67254223PRTC. necator 54Met Arg Arg Ile Glu Thr Ala Leu Leu Asp Gly Thr Trp Pro Pro Gly1 5 10 15Thr Arg Leu Pro Ala Glu Arg Val Leu Ala Gly Gln Tyr Glu Val Ala 20 25 30Arg Asn Thr Val Arg Glu Ala Ile Gln Arg Leu Ala Ala Arg Gly Leu 35 40 45Leu Gln Ser Arg Arg Gly Ala Gly Val Tyr Ala Thr Asp Gln Leu Arg 50 55 60Ala Gly Ile Ala Ser Pro Trp Gly Gln Leu Val Ala Asp His Pro Ala65 70 75 80Leu Arg Asp Asp Ile Leu Glu Phe Arg Arg Val Leu Glu Gly Ala Thr 85 90 95Ala Tyr Phe Ala Ala Leu Arg Ala Asp Ala Ala Asp Val Lys Arg Ile 100 105 110Arg Ala Leu Met Ala Glu Leu Glu Arg Ala Arg Ala Ala Asp Asp Lys 115 120 125Gln Ala Glu Ala Asp Ala Asp Ala Gln Leu His Asp Ala Ile Ala Gln 130 135 140Ala Ser His Asn Thr Met Phe Leu His Leu His Thr Ser Val Ile Gly145 150 155 160Met Leu Arg Glu His Ile Thr Ile Asn Gly Thr Gly Leu Arg Glu Gln 165 170 175Asp Asp Gly Ala Ser Asp Leu Leu Leu Leu Gln His Arg Thr Leu Cys 180 185 190Asp Ala Ile Cys Ala Arg Arg Pro Glu Glu Ala Arg Thr Ala Met Gln 195 200 205Thr His Ile Asp Phe Val Arg Ser Arg Val Glu Gln Asp Gly Ala 210 215 220551476DNAC. necator 55atgggcatgg tctcaataga acccggcagc tatcacgttg taccgccgca aaaccccggc 60gaggtcatcg atctgatccg gacgcacgcg atccaggtgg ttgacctcag gttcaccgac 120ttgcccggcg tgtggcagca cttctcgatc acgctgccgg aagtcaacga cgacttattc 180tctgtcggca tcggcttcga cgggtcttcc attcgcgggt tccaggaaat ccacgagtcc 240gacatgctgg tcaggcctga cccggccacg gcgttcatcg atccgtattg cgcagcacca 300acgctggcgc tgatctgcga cgtgctggac cctgtcctgc accagccgta ctcgcgcgat 360ccgcgccata tcgcacgcaa ggccgagctg tacctccggc aaaccggtct tgccacggtt 420tgctaccttg gccccgaact ggaattcttt attttcgact ccatacgctt cgggcaggac 480cagcactctg gctactacca tgtcgaatcc gccgaaggcg aatggacctc gggccgtgac 540gaaggcgcct atggcggggg caatctcggc tacaaacagc gctacaaggg cgggtacttt 600cccgtgccgc caagcgacac gctgcaggac atccgctccg aaatcgtgct cgcgctgatg 660caggccggca tccaggtcga agtgcatcac catgaggtcg ccacggccgg ccagaatgaa 720atcgacatgc gctttgcgcc actgatgcgc atggcagaca acgtgatgat gtacaagtac 780atctgcaaga acgttgcgcg ccgccacggc aaggtcgcca ccttcatgcc caagccgctg 840tttgccgaca acgcgagcgg catgcattgc caccagagcc tctggcgcga cgccgagaac 900ctcttctatg acaagaacgg ttgggcgcag acgtcccaga tgtgccgttg gtatataggc 960ggcttgctca gccacgcccc cgcgctgatg gcattctgcg caccgagcac gaactcctac 1020aagcggctgg tgcccggata cgaggcgcct gtcaatctgg ccatgtcgca gcgcaaccgg 1080tccgccgcgg cccggattcc aatggtttcg gattccccga gcgccaggcg cgttgaattt 1140cgctgtcctg atccgtcggc caatgcctac ctcgccttct cggcaatgct gctcgccggc 1200ctggacggca ttgaaaacca gacggacccc ggcgacccgc tcgacaagaa catctatgat 1260ctgccaccgg aggaagccgc ccgcatccgg caggtaccag gctccctgga agagtcgctt 1320tgcgcgctgg aagccgattc agcgttcctg cgcaaggggg atgtcttcac tgaagacctg 1380atcacgacct ggattgatta caagcgcacg cgcgagatcg atacattgaa ggtgcggcca 1440cacccgtggg aattccagct ctacttcgac atctga 147656491PRTC. necator 56Met Gly Met Val Ser Ile Glu Pro Gly Ser Tyr His Val Val Pro Pro1 5 10 15Gln Asn Pro Gly Glu Val Ile Asp Leu Ile Arg Thr His Ala Ile Gln 20 25 30Val Val Asp Leu Arg Phe Thr Asp Leu Pro Gly Val Trp Gln His Phe 35 40 45Ser Ile Thr Leu Pro Glu Val Asn Asp Asp Leu Phe Ser Val Gly Ile 50 55 60Gly Phe Asp Gly Ser Ser Ile Arg Gly Phe Gln Glu Ile His Glu Ser65 70 75 80Asp Met Leu Val Arg Pro Asp Pro Ala Thr Ala Phe Ile Asp Pro Tyr 85 90 95Cys Ala Ala Pro Thr Leu Ala Leu Ile Cys Asp Val Leu Asp Pro Val 100 105 110Leu His Gln Pro Tyr Ser Arg Asp Pro Arg His Ile Ala Arg Lys Ala 115 120 125Glu Leu Tyr Leu Arg Gln Thr Gly Leu Ala Thr Val Cys Tyr Leu Gly 130 135 140Pro Glu Leu Glu Phe Phe Ile Phe Asp Ser Ile Arg Phe Gly Gln Asp145 150 155 160Gln His Ser Gly Tyr Tyr His Val Glu Ser Ala Glu Gly Glu Trp Thr 165 170 175Ser Gly Arg Asp Glu Gly Ala Tyr Gly Gly Gly Asn Leu Gly Tyr Lys 180 185 190Gln Arg Tyr Lys Gly Gly Tyr Phe Pro Val Pro Pro Ser Asp Thr Leu 195 200 205Gln Asp Ile Arg Ser Glu Ile Val Leu Ala Leu Met Gln Ala Gly Ile 210 215 220Gln Val Glu Val His His His Glu Val Ala Thr Ala Gly Gln Asn Glu225 230 235 240Ile Asp Met Arg Phe Ala Pro Leu Met Arg Met Ala Asp Asn Val Met 245 250 255Met Tyr Lys Tyr Ile Cys Lys Asn Val Ala Arg Arg His Gly Lys Val 260 265 270Ala Thr Phe Met Pro Lys Pro Leu Phe Ala Asp Asn Ala Ser Gly Met 275 280 285His Cys His Gln Ser Leu Trp Arg Asp Ala Glu Asn Leu Phe Tyr Asp 290 295 300Lys Asn Gly Trp Ala Gln Thr Ser Gln Met Cys Arg Trp Tyr Ile Gly305 310 315 320Gly Leu Leu Ser His Ala Pro Ala Leu Met Ala Phe Cys Ala Pro Ser 325 330 335Thr Asn Ser Tyr Lys Arg Leu Val Pro Gly Tyr Glu Ala Pro Val Asn 340 345 350Leu Ala Met Ser Gln Arg Asn Arg Ser Ala Ala Ala Arg Ile Pro Met 355 360 365Val Ser Asp Ser Pro Ser Ala Arg Arg Val Glu Phe Arg Cys Pro Asp 370 375 380Pro Ser Ala Asn Ala Tyr Leu Ala Phe Ser Ala Met Leu Leu Ala Gly385 390 395 400Leu Asp Gly Ile Glu Asn Gln Thr Asp Pro Gly Asp Pro Leu Asp Lys 405 410 415Asn Ile Tyr Asp Leu Pro Pro Glu Glu Ala Ala Arg Ile Arg Gln Val 420 425 430Pro Gly Ser Leu Glu Glu Ser Leu Cys Ala Leu Glu Ala Asp Ser Ala 435 440 445Phe Leu Arg Lys Gly Asp Val Phe Thr Glu Asp Leu Ile Thr Thr Trp 450 455 460Ile Asp Tyr Lys Arg Thr Arg Glu Ile Asp Thr Leu Lys Val Arg Pro465 470 475 480His Pro Trp Glu Phe Gln Leu Tyr Phe Asp Ile 485 49057906DNAC. necator 57atgactgcgc ccttcgacat cctgacccgc atcgccgagc gcggccccgc gctgcgcctg 60gccgagcaga aggtggcgca ggtggtactg gaagacctgg ccggcgcggc cgccgccagc 120atcaatgagc tggcgcgcaa ggccggcgtc agcgaggcca gcgtgacgcg ctttgccaag 180gccatcggct gccgcgacgt gcgcgacctg aagctgcgcc tggcgcaggc caccgcggtg 240ggtgcgcgct tcctgcagcc cggcagcgtt cccgccggtg aagccacccc ggccacactc 300gctgacagca tccacgccga catcctcacc gcgctcgaag tcaaccgcgg catgatggat 360gcgcagcgca tcgaacaggc cgcgcgcctg ttgctcggcg cacgcatggt ctacgctttt 420ggcatgggcg gcggctcgtc gttcatggcc gacgaggccc gccatcgcct ggcgcgcctg 480ggccagccgg tggcgagcta ccaggacgcg ctgctacaga agatggtggc ggccacgctg 540ggccgcgacg acgtggtgct ggccttctcg gccagcggcc gcgtgccgga gatgctggcc 600agctgcgata tcgcgcgcga gtacggtgcc cgcctggtgg ccgtgaccgc gctgggctcg 660ccactggccg cgcgcgccga cgtgctgctg ccggtgcgca cgctggagac ggatttcatt 720ttcaaaccgt cggcatcgcg ctacgccatg ctgatggtgc tggacgtgct cgccacgcaa 780tgcgcgctgt tgcagccgga

ccagagcaaa gagcgcctgc gccggctcaa gtacgtgctg 840gacagccacc gcggcgaaag cggccccgcc cggggccctg acagccgcca gccgctcgga 900gactga 90658301PRTC. necator 58Met Thr Ala Pro Phe Asp Ile Leu Thr Arg Ile Ala Glu Arg Gly Pro1 5 10 15Ala Leu Arg Leu Ala Glu Gln Lys Val Ala Gln Val Val Leu Glu Asp 20 25 30Leu Ala Gly Ala Ala Ala Ala Ser Ile Asn Glu Leu Ala Arg Lys Ala 35 40 45Gly Val Ser Glu Ala Ser Val Thr Arg Phe Ala Lys Ala Ile Gly Cys 50 55 60Arg Asp Val Arg Asp Leu Lys Leu Arg Leu Ala Gln Ala Thr Ala Val65 70 75 80Gly Ala Arg Phe Leu Gln Pro Gly Ser Val Pro Ala Gly Glu Ala Thr 85 90 95Pro Ala Thr Leu Ala Asp Ser Ile His Ala Asp Ile Leu Thr Ala Leu 100 105 110Glu Val Asn Arg Gly Met Met Asp Ala Gln Arg Ile Glu Gln Ala Ala 115 120 125Arg Leu Leu Leu Gly Ala Arg Met Val Tyr Ala Phe Gly Met Gly Gly 130 135 140Gly Ser Ser Phe Met Ala Asp Glu Ala Arg His Arg Leu Ala Arg Leu145 150 155 160Gly Gln Pro Val Ala Ser Tyr Gln Asp Ala Leu Leu Gln Lys Met Val 165 170 175Ala Ala Thr Leu Gly Arg Asp Asp Val Val Leu Ala Phe Ser Ala Ser 180 185 190Gly Arg Val Pro Glu Met Leu Ala Ser Cys Asp Ile Ala Arg Glu Tyr 195 200 205Gly Ala Arg Leu Val Ala Val Thr Ala Leu Gly Ser Pro Leu Ala Ala 210 215 220Arg Ala Asp Val Leu Leu Pro Val Arg Thr Leu Glu Thr Asp Phe Ile225 230 235 240Phe Lys Pro Ser Ala Ser Arg Tyr Ala Met Leu Met Val Leu Asp Val 245 250 255Leu Ala Thr Gln Cys Ala Leu Leu Gln Pro Asp Gln Ser Lys Glu Arg 260 265 270Leu Arg Arg Leu Lys Tyr Val Leu Asp Ser His Arg Gly Glu Ser Gly 275 280 285Pro Ala Arg Gly Pro Asp Ser Arg Gln Pro Leu Gly Asp 290 295 300591344DNAC. necator 59gtgaattctc cctcccttga cgcattcctg gcgggggttg cccgccgcga ccccaatcaa 60cctgaattcc tccaggccgt gaaggaagtg atgatgacgc tctggccctt tgtcgagcgc 120aatccgcgct acgccgacca ggccctgctc gagcggctgg tggagcccga gcgcgtgatc 180cagttccgcg tggcctggac cgacgaccag aaccgggtgc aggtcaaccg cgccttccgc 240gtgcagcaca gctcggccat cggcccgttc aagggcggca tgcgcttcca cccgactgtg 300aacctgtcgg tgctgaagtt cctgggcttc gagcagacct tcaagaacgc gctgaccacg 360ctgcccatgg gcggcggcaa gggcggctcg gactttgatc ccaagggcaa gtccgatggc 420gaagtgatgc gtttctgcca ggcgctggtg accgagctgt tccgccacct gggcccggat 480accgacatcc cggccggcga catcggcgtg ggcgcacgtg aagtcggctt tatggccggc 540atgatgaaga agctttccaa ccagtccgcc tgcgtcttca ccggcaaggg cctggcctac 600ggcggcagcc tgatgcgccc ggaagcgacc ggctacggca cggtctactt tgcgcaggag 660atgctgcacc ggcgcgggcg cgctttcgac ggcctgcgcg tgctgatctc gggctcgggc 720aacgtggccc agtacgcggc cgagaaggcg atcgagctgg gcgccacggt gctgacgctg 780tccgattcag gcggcgtgct gcactacccg cagggcatga ctaccgagca gctggccgaa 840gtgatggcct tcaagaatga agagcgcggc cgcctgtctg actttgccgc ccgccacggc 900atggccttcg aagccggccg caccccgtgg cacgtgcccg ccgacgtggc gctgccgtgc 960gccacccaga acgagctgga cggcaacgac gccgagaccc tgctcggcaa tggcgtgatc 1020tgcgtggccg aaggcgccaa catgccgtcg acgctggaag ccgtggaccg ctttgtcgat 1080gcgaagatcc tctacgcccc gggcaaggcc agcaatgccg gcggcgttgc cacttccggc 1140ctggaaatgt cgcagaacgc catgcgcctg tcctggcacc atgccgaggt cgacgagaag 1200ctgcacgcga tcatgaagga catccaccag aactgcatcc accacgggca gaaggcggat 1260ggctatatca actacgtgga aggcgcgaac atcgccggct tcgtcaaggt agccgacgcc 1320atgctggcgc aaggcgtgat ctga 134460447PRTC. necator 60Met Asn Ser Pro Ser Leu Asp Ala Phe Leu Ala Gly Val Ala Arg Arg1 5 10 15Asp Pro Asn Gln Pro Glu Phe Leu Gln Ala Val Lys Glu Val Met Met 20 25 30Thr Leu Trp Pro Phe Val Glu Arg Asn Pro Arg Tyr Ala Asp Gln Ala 35 40 45Leu Leu Glu Arg Leu Val Glu Pro Glu Arg Val Ile Gln Phe Arg Val 50 55 60Ala Trp Thr Asp Asp Gln Asn Arg Val Gln Val Asn Arg Ala Phe Arg65 70 75 80Val Gln His Ser Ser Ala Ile Gly Pro Phe Lys Gly Gly Met Arg Phe 85 90 95His Pro Thr Val Asn Leu Ser Val Leu Lys Phe Leu Gly Phe Glu Gln 100 105 110Thr Phe Lys Asn Ala Leu Thr Thr Leu Pro Met Gly Gly Gly Lys Gly 115 120 125Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Asp Gly Glu Val Met Arg 130 135 140Phe Cys Gln Ala Leu Val Thr Glu Leu Phe Arg His Leu Gly Pro Asp145 150 155 160Thr Asp Ile Pro Ala Gly Asp Ile Gly Val Gly Ala Arg Glu Val Gly 165 170 175Phe Met Ala Gly Met Met Lys Lys Leu Ser Asn Gln Ser Ala Cys Val 180 185 190Phe Thr Gly Lys Gly Leu Ala Tyr Gly Gly Ser Leu Met Arg Pro Glu 195 200 205Ala Thr Gly Tyr Gly Thr Val Tyr Phe Ala Gln Glu Met Leu His Arg 210 215 220Arg Gly Arg Ala Phe Asp Gly Leu Arg Val Leu Ile Ser Gly Ser Gly225 230 235 240Asn Val Ala Gln Tyr Ala Ala Glu Lys Ala Ile Glu Leu Gly Ala Thr 245 250 255Val Leu Thr Leu Ser Asp Ser Gly Gly Val Leu His Tyr Pro Gln Gly 260 265 270Met Thr Thr Glu Gln Leu Ala Glu Val Met Ala Phe Lys Asn Glu Glu 275 280 285Arg Gly Arg Leu Ser Asp Phe Ala Ala Arg His Gly Met Ala Phe Glu 290 295 300Ala Gly Arg Thr Pro Trp His Val Pro Ala Asp Val Ala Leu Pro Cys305 310 315 320Ala Thr Gln Asn Glu Leu Asp Gly Asn Asp Ala Glu Thr Leu Leu Gly 325 330 335Asn Gly Val Ile Cys Val Ala Glu Gly Ala Asn Met Pro Ser Thr Leu 340 345 350Glu Ala Val Asp Arg Phe Val Asp Ala Lys Ile Leu Tyr Ala Pro Gly 355 360 365Lys Ala Ser Asn Ala Gly Gly Val Ala Thr Ser Gly Leu Glu Met Ser 370 375 380Gln Asn Ala Met Arg Leu Ser Trp His His Ala Glu Val Asp Glu Lys385 390 395 400Leu His Ala Ile Met Lys Asp Ile His Gln Asn Cys Ile His His Gly 405 410 415Gln Lys Ala Asp Gly Tyr Ile Asn Tyr Val Glu Gly Ala Asn Ile Ala 420 425 430Gly Phe Val Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile 435 440 445611395DNAC. necator 61atgaagtcag agttgagtgc gcatcttgcg gcgccagccg gggcgctgac cccggctccg 60cggttcaaca ccgtcgaaga cgctcaagac tatctgctgg cgcgcggcgt cagctacgtc 120ctcgcgcagt tcgtcgatat ccacggcgtc gccaaggcca aatcggtgcc agtggcgcat 180cttggctcgg tcctggcaga gggtgcgggc tttgccggct ttgccatctg gggcgtcggc 240attgaaccgc atggcccgga cttcatggct cgcggggatc tcgataccat cgggctggtg 300ccatggcagc cgggcctggc ccggatcgta tgcgaagggc atgtcgatgg tgcgccatgg 360cagtacgaca gccgggtcgt cctgaagcgg cagatcgcca ggctctcgca aggtggctac 420acgctttata ccggcctcga gcctgagttt tcgctgctgc gccgtgacga caaaggcggc 480atcggtccgt gcgacccgag cgatacgctg gccaagcctt gctacgacta caaggggctg 540tcgcgcacgc gtactttcct ggagcgtctg tccaacggac tgcgcgccgc gggaatcgat 600gtctaccaga tcgaccatga agacgccaac gggcaattcg aactgaatta caccttcacc 660gactgcctga cgtcttgcga ccatttcatc ttcttcaaga tggcggcatc ggaaatcgcc 720aacgagctgg gcctggtgtg ctccttcatg ccgaagccgt ttgcaaaccg ccccggcaac 780ggcatgcata tgcacatgtc gatcggcgat ggccagcgca acctgtttgc tgacaagagc 840gacccgcgcg ggctggacct gtcccagctg gcctatcact tccttggcgg gctgctggcc 900catgcgccgg cgctcacggc gctttgcgcg cccaccgtca attcctacaa gcggctcgtg 960gtgggccgtt ccctcaccgg cgccacctgg gctcccgcgt atatcagcta tggcgacaac 1020aatcgctcga gcatgatccg catccccaag ggccggctgg aactgcggct gcccgatggc 1080gccgccaacc cgtatctcgc cacagcggcc gtgattgcgg cggggcttga cggcatcgac 1140cggaagctcg atcccggcgc accgcgcaac accaatctct atgagtggag cgaggcgcag 1200ctggccgaag ccggcatcgg cctgctaccc cagaaccttg gcagcgcgct ggatgccctc 1260gaagccgatg cgctgataac cgaggcactg ggacccgttg cgcaagagtt cctgaagctc 1320aagcgcatgg agtggctcga atatcaacgg catgtctcgg actgggaagt gaagcagtac 1380ctggaattct tttga 139562464PRTC. necator 62Met Lys Ser Glu Leu Ser Ala His Leu Ala Ala Pro Ala Gly Ala Leu1 5 10 15Thr Pro Ala Pro Arg Phe Asn Thr Val Glu Asp Ala Gln Asp Tyr Leu 20 25 30Leu Ala Arg Gly Val Ser Tyr Val Leu Ala Gln Phe Val Asp Ile His 35 40 45Gly Val Ala Lys Ala Lys Ser Val Pro Val Ala His Leu Gly Ser Val 50 55 60Leu Ala Glu Gly Ala Gly Phe Ala Gly Phe Ala Ile Trp Gly Val Gly65 70 75 80Ile Glu Pro His Gly Pro Asp Phe Met Ala Arg Gly Asp Leu Asp Thr 85 90 95Ile Gly Leu Val Pro Trp Gln Pro Gly Leu Ala Arg Ile Val Cys Glu 100 105 110Gly His Val Asp Gly Ala Pro Trp Gln Tyr Asp Ser Arg Val Val Leu 115 120 125Lys Arg Gln Ile Ala Arg Leu Ser Gln Gly Gly Tyr Thr Leu Tyr Thr 130 135 140Gly Leu Glu Pro Glu Phe Ser Leu Leu Arg Arg Asp Asp Lys Gly Gly145 150 155 160Ile Gly Pro Cys Asp Pro Ser Asp Thr Leu Ala Lys Pro Cys Tyr Asp 165 170 175Tyr Lys Gly Leu Ser Arg Thr Arg Thr Phe Leu Glu Arg Leu Ser Asn 180 185 190Gly Leu Arg Ala Ala Gly Ile Asp Val Tyr Gln Ile Asp His Glu Asp 195 200 205Ala Asn Gly Gln Phe Glu Leu Asn Tyr Thr Phe Thr Asp Cys Leu Thr 210 215 220Ser Cys Asp His Phe Ile Phe Phe Lys Met Ala Ala Ser Glu Ile Ala225 230 235 240Asn Glu Leu Gly Leu Val Cys Ser Phe Met Pro Lys Pro Phe Ala Asn 245 250 255Arg Pro Gly Asn Gly Met His Met His Met Ser Ile Gly Asp Gly Gln 260 265 270Arg Asn Leu Phe Ala Asp Lys Ser Asp Pro Arg Gly Leu Asp Leu Ser 275 280 285Gln Leu Ala Tyr His Phe Leu Gly Gly Leu Leu Ala His Ala Pro Ala 290 295 300Leu Thr Ala Leu Cys Ala Pro Thr Val Asn Ser Tyr Lys Arg Leu Val305 310 315 320Val Gly Arg Ser Leu Thr Gly Ala Thr Trp Ala Pro Ala Tyr Ile Ser 325 330 335Tyr Gly Asp Asn Asn Arg Ser Ser Met Ile Arg Ile Pro Lys Gly Arg 340 345 350Leu Glu Leu Arg Leu Pro Asp Gly Ala Ala Asn Pro Tyr Leu Ala Thr 355 360 365Ala Ala Val Ile Ala Ala Gly Leu Asp Gly Ile Asp Arg Lys Leu Asp 370 375 380Pro Gly Ala Pro Arg Asn Thr Asn Leu Tyr Glu Trp Ser Glu Ala Gln385 390 395 400Leu Ala Glu Ala Gly Ile Gly Leu Leu Pro Gln Asn Leu Gly Ser Ala 405 410 415Leu Asp Ala Leu Glu Ala Asp Ala Leu Ile Thr Glu Ala Leu Gly Pro 420 425 430Val Ala Gln Glu Phe Leu Lys Leu Lys Arg Met Glu Trp Leu Glu Tyr 435 440 445Gln Arg His Val Ser Asp Trp Glu Val Lys Gln Tyr Leu Glu Phe Phe 450 455 460

Claims

1. A method for increasing carbon-based chemical product yield in an organism, said method comprising modulating activity of one or more polypeptides or functional fragments thereof involved in a stringent response and/or in a regulatory network of an organism selected from a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto, thereby increasing carbon-based chemical product yield in the organism as compared to an organism without said modulated polypeptide activity.

2. The method of claim 1 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene which limits intracellular glutamine, thereby inducing the stringent response.

3. The method of claim 2 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene selected from glutamate dehydrogenase (gdhA; H16_A0471, H16_B1945, or H16_A1356), glutamine synthetase (glnA; H16_A2335, H16_B0618 or H16_B2191) or glutamate synthase (g1tAB; H16_A3430 or H16_A3431).

4. The method of claim 2 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene selected from GlnB (H16_A0320 and H16_0750), NtrB (H16_A2333), NtrC (H16_A2332), PtsN (H16_A0384), GlnD (H16_A2057), spoT1 (H16_A0955) and/or spoT2 (H16_A1337).

5. The method of claim 2 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene selected from NtrC (H16_A2332), RpoN/sigma 54 (H16_A0387), sigma 54 modulation protein S30EA (H16_A0386), or homologs thereof.

6. The method of claim 1 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene effecting a signalling pathway leading to increased (p)ppGpp levels and induction of the stringent response in non-limiting culture conditions.

7. The method of claim 1 wherein the one or more polypeptides or functional fragments thereof is encoded by a gene involved in the regulatory network are selected from FadP (H16_A0459), PsrA (H16_A3736), LldR (H16_B0094), GlpR (H16_A2504), AccR/BphQ (H16_A1372 or H16_A1373) and/or HexR (H16_A1177 or H16_B1210) and/or are involved in a regulatory network involving glycerol, the Ntr system and/or a sucA riboswitch.

8. The method of claim 1, wherein modulating the activity of one or more polypeptides or functional fragments thereof comprises overexpressing or mutating an endogenous or exogenous nucleic acid sequence in the organism.

9. The method of claim 1, wherein modulating the activity of one or more polypeptides or functional fragments thereof comprises downregulating, deleting or mutating an endogenous or exogenous nucleic acid sequence in the organism.

10. A nonnaturally occurring organism capable of yielding a carbon-based chemical product, said organism selected from a species of Cupriavidus or Ralstonia with diminished polyhydroxybutyrate synthesis or an organism with properties similar thereto and comprising a modification to one or more polypeptides or functional fragments thereof encoded by a gene involved in a stringent response and/or in a regulatory network of the organism.

11. The nonnaturally occurring organism of claim 10 wherein modulating the one or more polypeptides or functional fragments thereof in the organism increases carbon-based chemical product yield in the organism as compared to an organism without said modulated polypeptide activity.

12. The nonnaturally occurring organism of claim 10 wherein the one or more modified polypeptides or functional fragments thereof are encoded by a gene which limits intracellular glutamine, thereby inducing the stringent response.

13. The nonnaturally occurring organism of claim 10 wherein the one or more polypeptides or functional fragments thereof are encoded by a gene or genes selected from glutamate dehydrogenase (gdhA; H16_A0471, H16_B1945, or H16_A1356), glutamine synthetase (ginA; H16_A2335, H16_B0618 or H16_B2191) or glutamate synthase (gltAB; H16_A3430 or H16_A3431).

14. The nonnaturally occurring organism of claim 10 wherein the one or more polypeptides or functional fragments thereof are encoded by a gene or genes selected from GlnB (H16_A0320 and H16_0750), NtrB (H16_A2333), NtrC (H16_A2332), PtsN (H16_A0384), GlnD (H16_A2057), spoT1 (H16_A0955) and/or spoT2 (H16_A1337).

15. The nonnaturally occurring organism of claim 10 wherein the one or more polypeptides or functional fragments thereof are encoded by a gene or genes selected from NtrC (H16_A2332), RpoN/sigma 54 (H16_A0387), sigma 54 modulation protein S30EA (H16_A0386), or homologs thereof.

16. The nonnaturally occurring organism of claim 10 wherein the one or more polypeptides or functional fragments thereof are encoded by a gene or genes effecting a signaling pathway leading to increased (p)ppGpp levels and induction of the stringent response in non-limiting culture conditions.

17. The nonnaturally occurring organism of claim 10 wherein the one or more polypeptides are encoded by genes involved in the regulatory network selected from FadP (H16_A0459), PsrA (H16_A3736), L1dR (H16_B0094), GlpR (H16_A2504), AccR/BphQ (H16_A1372 or H16_A1373) and/or HexR (H16_A1177 or H16_B1210) and/or are involved in a regulatory network involving glycerol, the Ntr system and/or a sucA riboswitch.

18. A method for producing a carbon-based chemical product in an organism, said method comprising fermenting the nonnaturally occurring organism of claim 10 with a biological or non-biological carbon source.

19. The method of claim 18 wherein modulating the one or more polypeptides or functional fragments thereof in the nonnaturally occurring organism increases carbon-based chemical product yield in the organism as compared to an organism without said modulated polypeptide activity.

20. The method of claim 18 wherein the one or more modified polypeptides or functional fragments thereof in the nonnaturally occurring organism are encoded by: a gene which limits intracellular glutamine, thereby inducing the stringent response; a gene or genes selected from glutamate dehydrogenase (gdhA; H16_A0471, H16_B1945, or H16_A1356), glutamine synthetase (ginA; H16_A2335, H16_B0618 or H1682191) or glutamate synthase (g1tAB; H16_A3430 or H16_A3431); a gene or genes selected from GlnB (H16_A0320 and H16_0750), NtrB (H16_A2333), NtrC (H16_A2332), PtsN (H16_A0384), GlnD (H16_A2057), spoT1 (H16_A0955) and/or spoT2 (H16_A1337); a gene or genes selected from NtrC (H16_A2332), RpoN/sigma 54 (H16_A0387), sigma 54 modulation protein S30EA (H16_A0386), or homologs thereof; a gene or genes effecting a signaling pathway leading to increased (p)ppGpp levels and induction of the stringent response in non-limiting culture conditions; and/or genes involved in the regulatory network selected from FadP (H16_A0459), PsrA (H16_A3736), LldR (H16_B0094), GlpR (H16_A2504), AccR/BphQ (H16_A1372 or H16_A1373) and/or HexR (H16_A1177 or H16_B1210) and/or are involved in a regulatory network involving glycerol, the Ntr system and/or a sucA riboswitch.